[The indicators of anti-endotoxin immunity under preeclampsia].

The dysregulation of immune mechanisms of control of gestational process results in a number of obstetric complications, including preeclampsia. Nowadays, there is no detailed description of characteristics of anti-endotoxin immune response under this complication. To evaluate the role of anti-endotoxin immunity in development of preeclampsia the determination of levels of LBP, IgG to core-region LPS, IgA, IgM, IgG in blood plasma was made in 134 pregnant women during II-III terms. The main group (74 pregnant women with preeclampsia) was divided on 2 sub-groups: 50 women with mild preeclampsia of and 24 women with severe preeclampsia. In the first sub-group, in 14 women with mild preeclampsia additionally concentration of soluble form of CD14 and BPI was determined. The control group consisted of 60 pregnant women with physiological course of gestation. The study demonstrated increase of level of LBP up to 14%, double increase of titers IgG to core-region LPS in blood plasma of pregnant women with preeclampsia under standard concentrations of IgA, IgM, IgG. It was demonstrated that under mild preeclampsia an increase of sCD14, BPI occurs as compared with standard values. The results of study reflect characteristic alterations of congenital and adaptive components of anti-endotoxin immunity under preeclampsia.

[Leading role of infection in formation of placental insufficiency].

AIM: Evaluate the role of endotoxinemia in pregnancy complicated with placental insufficiency. MATERIALS AND METHODS: Complex clinical-laboratory examination of 130 pregnant women in 70 of which placental insufficiency was diagnosed (main group) was carried out. The examinees of the main group were divided into 3 subgroups: 36, 20 and 14 pregnant women with compensated, subcompensated and decompensated placental insufficiency, respectively. The control group was composed of 60 pregnant women with physiological course of gestation. Levels of LPS, LPS-binding protein and IgG against core-region, cytokines (TNF-alpha, IL13, IL2, IL4, IL6, IL8, IL10, IFNgamma), C-reactive protein were determined in blood plasma. Infection by Chlamydia trachomatis, Mycoplasma genitalium, Treponema vaginalis, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum were evaluated by PCR. RESULTS: In the main group in 64.2% of cases the presence of genital tract infection was established, in 47.0%--urinary system. An increase of LPS level, titers of IgG against LPS core-region and LPS-binding protein in blood plasma of pregnant women with placental insufficiency was shown. Cytokine profile in placental dysfunction was characterized by a significant increase of IL1beta, IL8, TNF-alpha Th1 cytokine and IL10 Th2 cytokine concentrations and a decrease of pro-inflammatory IL2, IFNgamma levels. CONCLUSION: The results indicate a leading role of infection in formation of placental dysfunction as well as prove involvement of LPS Gram-negative bacteria in pathogenesis of this complication.

[The influence of the surgical access on the endotoxemia level].

The systemic inflammatory response, induced by the lipopolysaccharides of the normoflora of the operated patients was studied on the example of the plasma changes in 107 patients. All patients were divided in 4 groups, depending on the level of lipopolysaccharides in plasma. Thus, patients with the complicated course of the acute cholecystitis, demonstrated the increased lipopolysaccharides levels. The last correlates with the surgical access, the duration of the operation and early complications development. The results may be used for the diagnostics and decision making in emergency situations.

[Systemic endotoxinemia as a pathogenetic factor of pregnancy complication].

AIM: Evaluation of the role of hemocirculation of a bacterial endotoxin in pregnancy complicated with gestosis. MATERIALS AND METHODS: A complex clinical-laboratory examination of 74 pregnant women at the II-III trimester of gestational period including determination of serum cytokine levels (TNFalpha, IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IFNgamma), C-reactive protein, LPS, LPS-binding protein and IgG against R-core LPS in blood plasma was performed. RESULTS: In pregnant women with gestosis an increase in endotoxinemia level (p < 0.0001), twofold increase in IgG titers against R-core LPS (p < 0.01) and an increase in LBP concentration by 14% (p < 0.05) without an increase in concentration of C-reactive protein were shown. Cytokine profile in gestosis was characterized by a twofold statistically significant increase of IL-1beta (p < 0.0001), IL-8 (p < 0.0001) pro-inflammatory cytokine levels, a tendency of IL-6 (p = 0.063), TNFalpha (p = 0.13) increase and a decrease of Th1 cytokine concentrations: IL-2 - by 6.5 times (p < 0.0001), IFNgamma--by 2 times (p < 0.0001). CONCLUSION: The results allow to consider gestoses as a manifestation of increased endotoxin translocation into systemic blood flow during gestation, that determines the necessity to improve therapeutic measures in this pathology.

[The genetic markers of pathogenicity of S. enteritidis, antibiotic resistance of cultures and clinical characteristics of disease].

The article presents the results of evaluation of laboratory diagnostic value of relationships between the rate of detection of fragments of genes of pathogenicity islands associated with E. coli pathogenicity in clinical strains of S. enteritidis, antibiotic resistance of cultures and clinical characteristics of salmonellosis in adults. The sample included 105 patients with salmonellosis to apply the genetic test to isolated strains of S. enteritidis. The E. coli fragments of genes of pathogenicity islands are detected in 102 out of 105 analyzed strains of S. enteritidis (97.1%). The fragments specific to hlyA E. coli are detected in various qualitative and quantitative combinations in 50 (49.0%) clinical strains of S. enteritidis, hlyB--in 54 (52.9%), sfaG--in 57 (55.9%) and sfaA--in 29 (28.4%). The correlation is established between presence of fragments of genes of pathogenicity islands in S.enteritidis associated with E. coli pathogenicity and resistance to ampicillin, doxycycline, cephasoline, chloramphenicol, carbenicillin. The dependence of disease severity from S. enteridis poly-resistance to antibiotics is established.

[The comparative evaluation of informativity of methods of etiologic diagnostics of community-acquired pneumonia].

The article deals with the results of selection and testing of oligonucleotide primers to gene 16S pRN4 of a number of agents of community'-acquired pneumonia (Streptococcus pneumoniae. Haemorhilus influenzae. Moraxella catarrhalis, Klebsiella pneumonia. Pseudomonas aeruginosa. StaphyIococcus aureus) for their highly specified detection in clinical material on application of polymerase chain reaction. The comparison with bacteriological method (golden standard) is used to demonstrate the high sensibility, specificity and diagnostic effectiveness of product enabling to apply it in practice to diagnose communitly-acquired pneumonia regardless of duration, severity and phase of disease and to develop corresponding diagnostic systems.

[Character of lipopolysaccharide-binding protein level changes in different pathological conditions and dysbiosis].

AIM: To evaluate diagnostic value of lipopolysaccharide-binding protein (LBP) level in different infectious processes and dysbiosis. MATERIALS AND METHODS: Serum samples of patients with salmonellosis, urogenital chlamydiosis, community-acquired pneumonia, polypous rhinosinusitis, and bacterial vaginosis were studied. RESULTS: LBP level were lower in patients with salmonellosis compared to healthy persons and decreased with increasing severity of the disease. Higher levels of LBP were detected in patients with chlamydiosis. Direct correlation between LBP level and etiology and severity of community-acquired pneumonia was demonstrated. In patients with polypous sinusitis, LBP level correlated directly with duration of disease, and inversely--with duration of remission. Twofold increase of mean LBP blood concentration and its correlation with duration of dysbiosis in patients with bacterial vaginosis were revealed. CONCLUSION: Multidirectional modulation of antiendotoxin defense was observed in different pathological conditions.

[Development and use of test-systems for molecular-genetic detection of out-of-hospital pneumonia agents].

AIM: Search of new targets and improvements of molecular-genetic detection methods of species specific DNA fragments of out-of-hospital pneumonia (OP) infection agents in sputum. MATERIALS AND METHODS: Species specific oligonucleotide primers to Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Branhamella catarrhalis were selected. 103 sputum samples from OP patients and 15 from practically healthy humans were test by using PCR and bacteriological method. RESULTS: Mixed bacterial flora was detected by bacteriologic method in OP in 26 cases (25.2%). 14 patients (13.6%) had B. catarrhalis DNA in sputum, 12 patients (11.6%)--S. pneumoniae, H. influenzae was detected in 11 examined individuals (10.7%), K. pneumoniae (7.9%)-- in 8. In 32 cases (30%) etiology of OP could not be deciphered bacteriologically. B. catarrhalis was detected in 19 (18.5%) of OP patients, S. pneumoniae--in 17 (16.5%), H. influenzae and K. pneumoniae--in 14.5% and 12.6% (15 and 13 patients) respectively by Using the developed PCR primers. In 16.5% cases (17 patients) positive results against several bacteria species simultaneously were obtained by PCR method, in 3.9% (4 patients) P. aeruginosa was detected. CONCLUSION. Use of PCR for detection of out-of-hospital pneumonia agents DNA significantly increases informativity of the study when compared with bacteriological method.

[Genetic mechanisms of Salmonella enteritidis biodiversity and clinical features of salmonellosis].

AIM: To assess prevalence of fragments of Escherichia coli pathogenicity islands in Salmonella enteritidis strains as well as to study clinical signs of disease caused by these strains in adults. MATERIALS AND METHODS: Ninety-six patients with salmonellosis were studied. Ninety strains of S. enteritidis were isolated and tested by PCR for the presence of genes associated with pathogenicity islands of E. coli: hlyA, hlyB, sfaG, and sfaA. RESULTS: It was determined that DNA fragments homologous to pathogenicity islands of E. coli were present in 87 (96.7%) of S. enteritidis clinical isolates. Disease caused by Salmonella strains which possess only sfaG was mostly mild--7 (33.3%), whereas strains which had sfaG with fragments of hlyA and/or hlyB caused severe disease--7 (50%). sfaA fragments were found mostly in combination with other genes. In such cases the disease was mostly severe--6 (42.8%). CONCLUSION: Correlation between presence of E. coli pathogenicity islands in Salmonella spp., their antibiotic resistance and severity of infection was established.

[State of antiendotoxin defense during community-acquired pneumonia].

AIM: To assess the effect of systemic endotoxinemia and LPS-induced immune response on severity of community-acquired pneumonia (CAP). MATERIALS AND METHODS: Clinico-immunological follow-up of 60 patients with CAP of different severity was conducted. LPS was quantitatively measured by ENDOSAFE ENDOCHROME (0.015-0.12 EU/ml), LBP--by Hbt Human LBP ELISA (min 1 ng/ml), IgG to core region of LPS--by Hbt EndoCAb ELISA (min 0.125 MU/ml) ("HyCult biotechnology", Netherlands). RESULTS: Level of LPS in blood was not increased, whereas there was increase of antiendotoxin proteins in patients with more severe CAP with Gram-negative or mixed bacterial flora identified. Correlation between levels of LBP, IgG to core-region of LPS, IL-6, TNF-alpha, IL-4 and immune response abnormalities during severe forms of disease was demonstrated. CONCLUSION: State of antiendotoxin immunity is an etiologic marker of CAP and its severity.

[Procedure for Trichomonas vaginalis cultivation].

The investigation was undertaken to design a nutrient medium that could reduce the duration of a culture test in the diagnosis of trichomoniasis and to comparatively evaluate its effectiveness. A procedure was proposed to cultivate Trichomonas vaginalis, which was based on the use of the nutrient medium containing casein hydrolysate, yeast autolysate, and serum, added by 0.6 ml of liquid Ferrum Lec (iron (III) hydroxide polyisomaltose), which reduced the time taken to identify Trichomonas vaginalis (on day 2). At the same time, the higher content of iron [III] hydroxide with polyisomaltose was shown to produce no positive effect.

[Endotoxinemia and anti-endotoxin immunity in women with bacterial vaginosis].

AIM: To assess the level of endotoxinemia and state of antiendotoxin immunity compared with parameters of humoral immunity in patients with bacterial vaginosis. MATERIALS AND METHODS: Blood sera from 28 women with bacterial vaginosis (main group) and 24 clinically healthy women (control group). Concentration of lypopolysaccharide (LPS) in sera was measured by semi-quantitative gel-clot test using LAL-reagent Endosafe. Concentration of LBP, IgG EndoCAb as well as levels of serum IgA, IgM and IgG were measured by enzyme immunoassay. RESULTS: Conditionally higher level of LPS, 2-fold increase of LBP concentration, 1.7-fold increase of IgG EndoCAb as well as tendency to decrease of IgA level were revealed in sera of women from the main group. Dysbiotic shift in vaginal microecological system of non-pregnant women of child-bearing age was associated with breakthrough of LPS in systemic bloodstream. Registration of vaginosis during pregnancy associated with detection of Chlamydia, possessing LPS similar to enterobacteria, revealed presence of unfavorable variants of prolonged torpid course of dysbiotic signs. CONCLUSION: During torpid forms of bacterial vaginosis, entotoxinemia and abnormalities in anti-endotoxin immunity are revealed, which the need of optimization of its therapy.

[Preparation glycosaminosulphate efficacy in comprehensive treatment of patients with odontogenic phlegmon of maxillofacial region].

The results of 20 patients (with odontogenic phlegmon of maxillofacial region) treatment by glycosaminosulphate were compared with the results of patients received traditional treatment. Significant speeding-up of the blood indices normalization terms, humoral immunity factors and postoperative wound regeneration after odontogenic phlegmon of maxillofacial region lancing was determined.

[Proinflammatory cytokines in children with acute enteric infections caused by enterobacteria].

The levels of the proinflammatory cytokines including interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) was investigated in 73 children in the age range from 3 to 17 years with slight and medium-heavy diarrheal illness caused by pathogenic and conditionally pathogenic entrobacteria. Strong positive correlation was found between: the levels of IL-1beta, TNF-alpha and intoxication rate (r=0.65 and r=0.49); height of temperature and duration of fever (r=0.86 and r=0.66); dyspepsia (r=0.48 and r=0.41); diarrhea (r=0.37 and r=0.54) and changes in blood including number of leucocytes (r=0.40 and r=0.52) and level of erythrocyte sedimentation rate (ESR) (r=0.65 and r=0.52). Medium positive relationship between the level of IL-6 and intoxication rate (r=0.40 and r=0.52), height of temperature and duration of fever (r=0.45), changes in blood including ESR (r=0.42) and number of leucocytes (r=0.46) was shown. There was a strong positive correlation between IL-1beta and TNF-alpha (r=0.74), between IL-6 and TNF-alpha (r=0.71) and a medium positive correlation between IL-1beta and IL-6 (r=0.61). Considerable decrease in concentration of all cytokines during early period of convalescent at disease with no complications was revealed. Change in concentration of IL-1beta and TNF-alpha determines both intoxication rate and fever. High level of IL-6 is related with complications and lingering course of disease.

[Pathogenicity factors of opportunistic enterobacteria and its role in development of diarrhea].

The data of pathogenicity factors of opportunistic enterobacteria, including Klebsiella, Enterobacter, Citrobacter, Proteus, Providencia and Hafnia species are submitted. The genetic control and a role of pathogenicity factors of opportunistic enterobacteria in development of diarrhea syndrome are presented. Data about adhesins, hemolysins, cytotoxic necrotizing factors and bacterial modulins are described. The characteristic of cytotonic and cytotoxic enterotoxins, including LT, ST, Shiga-like and cytolethal toxins, and mechanisms of diarrheagenic action are analysed. The role of bacterial lipopolysaccharide (endotoxin) and induction of locally synthesized proinflammatory cytokins in pathogenisis of diarrhea are discussed.

[Genetic markers of pathogenicity of conditionally pathogenic enterobacteria isolated from children with diarrheal diseases].

In the process of examination of 89 children from different age groups with diarrheal disease caused by bacteria from Enterobacteriaceae family 89 microorganisms were isolated including Klebsiella spp. (37 isolates), Citrobacter spp. (9 isolates), Enterobacter spp. (17 isolates), Hafnia alvei (1 isolate), Morganella morganii (11 isolates), Proteus spp. (14 isolates). Presence of genes associated with pathogenicity islands (PAIS): hlyA, hlyB (hemolysin), sfaG (fimbria antigen type S), cnf1 (cytotoxic necrotizing factor 1), estB (heat-stable enterotoxin B)were studied in these cultures by PCR. It was found that 32.6% of examined isolates had fragments of PAIS's genes--hlyA was detected in 9 cases (10.1%), hlyB--in 10 cases (11.2%), sfaG --in 8 cases (9%), cnf1--in 9 cases (10.1%), and estB--in 3 cases (3.4%). Positive correlation between genetic determinants hlyB and cnf1 as well as hlyA and sfaG was found while estB was not associated with other genes. Weak positive correlation between presence of sfaG and resistance to tetracycline and chloramphenicol was detected. Factors coded by revealed determinants of PAIS can play a role in the development of diarrheal syndrome.

[Bacterial vaginosis: etiopathogenetic aspects].

Current microbiological data on the vaginal microecology in healthy women and in patients with bacterial vaginosis are presented in the article. Problems of formation of colonization resistance of vaginal microbiocenosis were discussed. Etiologic role of certain microorganisms and their associations in the pathogenesis of bacterial vaginosis was considered. Pathophysiological processes leading to development of vaginal disbiosis, features of local immune status and molecular specificity of intercellular interactions in the development of adaptive immune response were characterized.

[Genetic determinants of Escherichia coli pathogenicity isolated from urine and feces of children with different clinical variants of urinary system infection]. / Geneticheskie determinanty patogennosti Escherichia coli, izolirovannykh iz mochi i fekalii detei s razlichnymi klinicheskimi variantami infektsii mochevoi sistemy.

In the process of examination of 156 children of different age groups 176 E. coli cultures were isolated; of these, 98 cultures were isolated from acute cystitis and pyelonephritis patients, 28--from urine in cases of aysmptomatic bacteriuria, 30--from feces in cases of asymtomatc bacteriuria and intestinal dysbacteriosis, while 20 cultures--from feces of healthy children. In these bacteria the presence of genes associated with pathogenicity islets (PI) hlyA, hlyB, cnf-1, papC, sfaG and gene irp-2 (iron-regulated protein) was established with PCR. The detection rate of PI determinants in uropathogenic E. coli (UPEC) was shown to depend on the variants of the clinical manifestation of urinary tract infection. The total detection rate of PI gene fragments in UPEC cultures of different origin was indicative of their definitely less frequent occurrence in asymptomatic bacteriuria, observed simultaneously with intestinal dysbacteriosis, in comparison with acute urological infection. Practically the same detection rate of PI determinants in E. coli, isolated in asymptomatic bacteriuria in children, reflected high probability of genetic exchange in the above-mentioned fragments and made it possible to presume the existence of DNA sites, characteristic mainly of pathogenic clones. The established heterogeneity of the detection rate of PI determinants in E. coli clinical isolates requires further study.