Formation of biofilms in human biotope microbiocenosis: model for prognostic calculations of intermicrobial relationships

Comparative ranging analysis of mixed microbial biofilm forming by microbes from the same biotope is performed on the example of lactobacilli and yeast-like fungi (Candida species). Lactobacillus species (L. acidophilus, L. casei) leader strains influencing yeast-like fungal specie-depended biofilms were established. L. acidophilus and L. casei leader strains completed each other and revealed opposite regulating properties in respect of biofilm forming by С. albicans and C. tropicalis strains. In comparison to unity of positions C. tropicalis strains (all strains as the unite block in ranging sequence) according to their biofilm forming, two С. albicans subspecie populations (two blocks with higher and lower biofilm forming) were identified in biofilm ranging sequences. Results indicate perspectivity of ranging biofilm forming (by coupled cultures of potentially antagonistic microorganisms

CO-FUNCTIONING OF LECTINS OF MULTICOMPONENT PROBIOTIC AND POTENTIAL PROBIOTIC COMPARTMENT OF BIOTOPE ON THE EXAMPLE OF AUTOREGULATOR LACTOBACILLUS SYSTEM

Using lectins of Acilact, clinical isolates of Lactobacillus from vaginal biotope were screened. Lactobacilli of the biotopes were classified to functional groups. Results indicate autoregulation signal properties of probiotic microbial lectins

Mobile synbiotope microbiocenosis against pathogens

Based on own results, conception of mobile synbiotope microcenosis against pathogens as well as new antimicrobial strategies connected to conception are proposed. Synbiotope mobility is determined by the presence (in varying combinations] of cofunctioning antimicrobial ways which can be realized with participation of highly molecular mass metabolites of probiotic bacterial cultures. Thus, probiotic enzymatic systems (proteinases, EPS-depolymerases and/ or oxidoreductases] reveal cofunctioning with probiotic lectins. New antipathogen strategies use technological advantages of choice and selection of symbiotic strains and consortia producing synergistical enzyme-lectin-glycoconjugate systems against microfungal and Gram-positive pathogens. Additional strategies involve construction of synbiotic systems supporting probiotic compartment of biotope

РОЛЬ ПОЛИМОРФИЗМОВ ГЕНА СRP В ВОЗНИКНОВЕНИИ ФИБРИЛЛЯЦИИ ПРЕДСЕРДИЙ У ПАЦИЕНТОВ ИБС

The purpose. Assessment of the contribution of CRP gene polymorphisms rs3093077, rs1130864 and rs1205 to the development of atrial fibrillation (AF) in CAD patients depending on gender and age.Material and methods. The study included 302 patients with stable coronary artery disease. C-reactive protein (CRP) concentration was measured by high-sensitive immunoturbidimetric assay. The genotyping was performed in 96-well plates using TaqMan assay.Results. The elderly age > 65 years and the presence of type 2 diabetes mellitus are the clinical and anamnestic predictors of AF development (р=0.003; 0.01). G/A rs1130864 CRP heterozygous carriers have a 4-fold increased risk of AF development among the patients with coronary artery disease (p=0.0025) according to the overdominant inheritance model. Men who are the carriers of С/Т rs1205 CRP genotype have on average a 5-fold increased risk of AF development in case of CAD presence (according to the overdominant model, р=0.022). A 5-fold increased risk of AF development is associated with men carrying G/A rs1130864 CRP genotype (p=0.015), meanwhile the men < 65 years, who carry the same genotype (rs1130864 CRP), have a 9-fold increased risk of AF development (p=0.025), as compared to the patients who are the carriers of other genotypes.The patients of both genders < 65 years, carrying the С/Т rs1205 CRP genotype, have a 10-fold increased risk of AF development as compared to the carriers of other genotypes (OR =10.03 (1.26-79.59); 95% CI).Conclusion. The genetic markers should be considered to assess the risk of AF development. Цель. Оценить вклад полиморфизмов гена CRP rs3093077, rs1130864, rs1205 в развитие фибрилляции предсердий (ФП) у пациентов ИБС в зависимости от пола и возраста.Материалы и методы. В исследование включено 302 пациента со стабильной ИБС. Концентрацию hs-СРБ определяли с помощью высокочувствительного иммунотурбидиметрического метода. Генотипирование осуществляли в 96-луночном формате методом TaqMan.Результаты. Клинико-анамнестическими предикторами возникновения ФП являются пожилой возраст > 65 лет и наличие у пациента СД 2-го типа (р = 0,003; 0,01). Носительство гетерозиготного генотипа G/A rs1130864 CRP увеличивает риск развития ФП у пациентов с ИБС более чем в четыре раза (р=0.0025) по сверхдоминантной модели наследования. У мужчин, носителей генотипа С/Т rs1205 CRP, риск развития ФП при ИБС увеличивается в среднем в пять раз (сверхдоминантная модель наследования (р=0,022). У мужчин, носителей генотипа G/A rs1130864 CRP, определяется увеличение риска развития ФП в 5 раз (р=0,015), а у мужчин в возрасте < 65 лет, носителей того же генотипа rs1130864 CRP, риск развития ФП увеличивается в 9 раз (р=0,025), в отличие от пациентов, носителей других генотипов. У пациентов обоих полов < 65 лет, носителей генотипа С/Т rs1205 CRP, риск развития ФП в 10 раз выше, чем у носителей других генотипов (ОШ=10,03 (1.26-79.59); 95% ДИ.Заключение. Для оценки риска возникновения ФП необходимо учитывать генетические маркеры.

Состояние легочной микробиоты у мышей С57bl/6 в модели экспериментального туберкулеза

The objective: to study the changes in the lung microbiota in inbred C57BL/6 mice after aerogenic infection with M. tuberculosis in an experimental tuberculosis model.Subjects and Methods. This study was carried out on 20 female mice of inbred line C57BL/6 weighing 20-22 grams which were infected in a Glas-Col aerosol chamber (USA) with the culture of M. tuberculosis of virulent strain H37Rv at the dose of 400 CFU/lung. Morphological and microbiological assessment of the lungs state was performed before (day 0) (n=5) and 7 (n=5), 30 (n=5) and 60 (n=5) days after the infection. The results obtained were subjected to statistical processing using ANOVA test and Student t-test.Results. 7, 30, and 60 days after aerosol infection with M. tuberculosis against the background of successive morphological and microbiological changes typical of the experimental tuberculosis model, we established an imbalance of bacterial population in the lung microbiota. Before infection with M. tuberculosis, a scanty biotope was recorded with a predominance of lactobacilli –Lactobacillus murinus, Lactobacillus apodeme. 7, 30 and 60 days after infection with M. tuberculosis, consistent changes were recorded, such as increase in the number and diversity of the bacterial population. The most indicative markers of the recorded imbalance were: Streptococcus thoraltensis, Streptococcus acidominiminus, Arthrobacter crystallopoietes, Staphylococcus hominis, Micrococcus luteus.Conclusion. Tuberculosis infection is a significant factor affecting the state of the lung microbiota. With increased duration of the infection with M. tuberculosis, imbalance of the bacterial flora is formed in the lungs of C57BL/6 mice, accompanied by characteristic tissue inflammation and growing mycobacterial load.Цель исследования: изучить динамику изменений в микробиоте легких у инбредных мышей C57BL/6 после аэрогенного инфицирования M. tuberculosis в модели экспериментального туберкулеза.Материалы и методы. Настоящая работа выполнена на 20 самках мышей инбредной линии C57BL/6, массой 20–22 грамм, которые были заражены в аэрозольной камере Glas-Col (США) культурой Мtb вирулентного штамма H37Rv в дозе 400 КОЕ/легкое. Морфологическую и микробиологическую оценку состояния легких проводили до (день 0) (n=5) и через 7 (n=5), 30 (n=5) и 60 (n=5) дней после инфицирования. Полученные результаты подвергали статистической обработке с использованием теста ANOVA и Стьюдент t-теста.Результаты. Через 7, 30 и 60 дней после аэрозольного инфицирования M. tuberculosis, на фоне последовательных морфологических и микробиологических изменений, характерных для модели экспериментального туберкулеза, нами установлен дисбаланс бактериальной флоры в микробиоте легких. До заражения Mtb регистрировали скудный биотоп с преобладанием лактобацилл – Lactobacillus murinus, Lactobacillus apodeme. Через 7, 30 и 60 дней после инфицирования Mtb регистрировали последовательные изменения в виде увеличения количества и разнообразия бактериального сообщества. Наиболее показательными маркерами регистрируемого дисбаланса были: Streptococcus thoraltensis, Streptococcus acidominiminus, Arthrobacter crystallopoietes, Staphylococcus hominis, Micrococcus luteus.Заключение. Туберкулезная инфекция является значимым фактором, воздействующим на состояние микробиоты легких.С увеличением длительности инфицирования Mtb в легких мышей C57BL/6 формируется дисбаланс бактериальной флоры, сопровождающийся характерным тканевым воспалением и нарастанием микобактериальной нагрузки

The relationship of gene polymorphisms of C-reactive protein with the development of myocardial infarction and formation of multifocal atherosclerosis in CHD patients

Purpose. To assess the contribution of the CRP gene polymorphisms rs3093077, rs1130864 and rs1205 in the development of myocardial infarction (MI) and multivessel disease (MVD) in CAD patients based on gender and age. Material and methods. 303 patients with stable coronary artery disease were included in the study. C-reactive protein (CRP) levels were measured by high sensitive immunoturbidimetric assay. The genotyping studies were performed in 96-well plates using the TaqMan assay. Results. Male gender and older age are proven to be among independent clinical predictors of myocardial infarction and the development of MVD in patients with coronary artery disease. The rs1205 C/C CRP homozygous carriers have a significantly higher risk of multivessel coronary lesions at age > 65, regardless of gender (OR = 4.72, 95 % CI = 1.27–17.56; p = 0.045). The C/C genotype of rs3093077, A/G of rs1130864 and P/T of rs1205 in female patients reduce the risk of myocardial infarction (OR = 0.53, 95 % CI = 0.30–0.95; p = 0.0079, OR = 0.37, 95 % CI = 0.16 – 0.82; p = 0.0027 and OR = 0.35, 95 % CI = 0.14–0.84; p = 0.0097, respectively). The groups with and without myocardial infarction were comparable by the allele frequencies and genotype distribution combinations of the CRP polymorphisms (rs3093077, rs1130864 and rs1205). Regardless of a positive history of MVD and myocardial infarction, serum levels of CRP over 3 mg/L indicates a high risk of cardiovascular events in patients with stable coronary artery disease. However, there is no relationship between protein levels and the polymorphisms of the genes encoding them (p = 0.56). Conclusion: The prediction of myocardial infarction and MVD requires not only assessing clinical and demographic data of patients, but also measuring CRP levels and studying its gene polymorphisms

Candida markers of urogenital biotopes diseases: reactivity to lectins of probiotics

Possibilities of using eukaryotic microorganisms (Candida spp.) of microbiocenoses of mucosal open cavities as potential biomarkers of pathological states of biotopes were investigated. To identify biotopes characterizing Candida strains we used new promising natural glycoconjugate recognition agents -lectins that imitate the effect of probiotics. The lectins were isolated from cultures of probiotic strains of lactobacillus and bifidus bacteria characterizing and supporting probiotic compartment of microbiocenosis of human mucosal open cavities as part of endogen sensory system "lectins of probiotic microorganisms - opportunistic eukaryotic yeast-like fungi". The study was carried out on examples of cases of dysbiotic disorders in the composition of microbiocenoses against the background of pathologies of the urogenital tract of patients with established diagnosis. Results demonstrate the early (before development of visible inflammatory reactions) presence of changed Candida cells in pathological biotopes of urogenital tract indicating states of diseases characterizing with increased differentiating affinities and sensitivities of Candida to a panel of probiotic lectins. Probiotic lectins are perspective in combinative prognostics and diagnostics of pathological states of functional biotopes of human mucosal open cavities as well as for prophylaxis and therapy of such states by compensation of deficit or absence of probiotic microflora to support healthy balance of mucosal immunity, instead of probiotics or in combinations with probiotics, antibiotics and other antimicrobial agents, therapeutics, and factors. The data also perspective in study of pathological biotope states possessing risk of conversion into tumor-like landscapes

LUNG DIFFUSION CAPACITY AND NON-SPECIFIC INFLAMMATION MARKERS IN PATIENTS WITH CORONARY HEART DISEASE AND DIABETES MELLITUS

Aim. To assess the differences in the lung diffusion capacity and non-specific inflammation markers among patients with coronary heart disease (CHD) and the combination of CHD and Type 2 diabetes mellitus (DM-2). Material and methods. The study included 70 CHD patients: with DM-2 (n=40; 57,1%) and without DM-2 (n=30; 42,9%). Lung diffusion capacity (DIco) was assessed with the single-breath method and subsequent correction by haemoglobin levels (DIco cor). The levels of the following non-specific inflammation markers were also assessed: IL-

THE RISK FACTORS AND EVALUATION CRITERIA FOR PROGRESSION OF ATHEROSCLEROSIS IN ONE YEAR POST CORONARY BYPASS

Aim. To assess risk factors and to compare evaluation criteria of non-coronary atherosclerosis progression in coronary heart disease patients in 1 year post coronary bypass surgery (CBG).Material and methods. Of 732 consequtive patients (586 males, 146 females, median age 59 y.o.) underwent CBG, 504 were investigated second time in 1 year after operation. Patients underwent ultrasound study (US) of carotid and peripheral arteries with measurement of intima-media thickness. Ankle-brachial index (ABI) was measured automatically with sphygmomanometer VaSeraVS-1000 (Fukuda Denshi,Japan). In blood, lipids were tested. Based on the stenosis indexes NASCET and ECST, the groups were selected: <30% (minor stenosis); 30-49% (moderate stenosis); 50-69% (severe stenosis); 70-99% (critical stenosis); occlusion; absence of stenoses. Criteria for progression of atherosclerosis was existence of at least one sign: transition of non-coronary stenosis from one group to another by US; decrease of ABI during 1 year below0,9 in normal baseline;  decrease of ABI >10% if the baseline ABI lower than 0,9. For analysis, two groups were selected: group 1 (n=375) without progression, and group 2 (n=129) with progression of atherosclerosis. The relation of possible factors to probaility of  on-coronary atherosclerosis progression was evaluated in logistic regression model.Results. Levels of the total cholesterol (TC) and low density lipoproteides cholesterol (LDL-C) were higher in patients with progression of atherosclerosis. In dynamics, significantly in both groups the level of high density lipoproteides cholestrol (HDL-C) increased (p<0,001), and triglycerides decreased (p<0,05). In intergroup analysis, ABI values were higher in the group with no progression of atherosclerosis (p<0,001). In dynamics, in the group of atherosclerosis progression there was significant ABI decrease (p<0,05). In non-progression group ABI, remaning in reference range, significantly decreased during a year after CBG (p<0,05). Thickness of CIM in the non-progression group significantly decreased during a year post-surgery, but in progression group the difference was non-significant. In monofactorial logistic regression, probability of atherosclerosis progression increased with the increase of age, in smoking anamnesis before surgery, in increase of TC and LDL-C, as with GFR decline and in baseline multifocal atherosclerosis presence. Multifactorial analysis showed remaining statistical significance for the several arterial regions, and adverse lipid profile.Conclusion. In one year post-CBG, progression of non-coronary atheroslerosis is marked in 25,6% of patients. In one year post-CBG, ABI decreases independently from the presence of non-coronary atherosclerosis. Factors associated with atherosclerosis progression, were multifocality, level of TC and LDL-C

THE ROLE OF C-REACTIVE PROTEIN AND POLYMORPHISMS OF ITS CANDIDATE GENES IN THE DEVELOPMENT OF IN-HOSPITAL CARDIOVASCULAR COMPLICATIONS IN PATIENTS AFTER CORONARY BYPASS SURGERY

Aim. To assess the prognostic significance of preoperation C-r.p. concentration and polymorphic site of the gene CRP (rs3093077, rs1130864, rs1205) for the development of early cardiovascular complications after direct myocardial revascularization.Material and methods. Totally 249 patients studied with CHD, underwent coronary bypass surgery (CBG). Concentration of C-r.p. was assessed with high-sensitive immune-turbidimetric method, genotyping was done by 96-hole format via TaqMan method, before CBG.Results. The risk of perioperational CVC increased in presence of such factors as age more than 65 y. o. (р=0,037), preoperational C-r.p. concentration higher than 5 mg/mL (р=0,026), homozygous genotype GG in promoter region of CRP gene (rs3093077) (c2 =9,08, р=0,0011) within all other conditions different (presence or absence of atrial fibrillation (AF), diabetes mellitus (DM) 2 type, duration of hypertension anamnesis (AH). In patients older than 65 y. o. the CVC risk increased almost 3 times: OR=2,8 (95% CI=1,07-7,34), and in serum concentrations of C-r.p. more than 5 mg/mL — two and a half times: OR=2,5 (95% CI=1,11-5,77). The carriage of genotype GG rs3093077 СRP increases the risk of CVC in in-hospital period of CBG for more than 2 times.Conclusion. For prediction of CVC in CBG it is necessary to evaluate not only clinical and anamnestic characteristics of a patient, but also the level of preoperation CRP, and genetic polymorphisms of their genes