[ANTIBIOTICS SENSITIVITY OF STAPHYLOCOCCUS AUREUS, PSEUDOMONAS AERUGINOSA AND BACTERIA OF BURKHOLDERIA CEPACIA COMPLEX, PERSISTING IN LUNGS OF PATIENTS WITH MUCOVISCIDOSIS (MV)].

AIM: Study the spectrum of resistance to antibiotics and its variability of Staphylococcus aureus, Pseudomonas aeruginosa and Burkholderia cepacia complex (BCC), persisting in lungs of MV patients. MATERIALS AND METHODS: 312 strains of S. aureus, 213 strains of P. aeruginosa, 186 strains of BCC were studied. Monitoring of antibiotics sensitivity was carried out in strains, isolated from 30 patients with chronic S. aureus infection, from 22 patients with chronic BCC infection and from 21 patients with chronic pseudomonas infection. Interval of monitoring was from 14 days to 5 years 7 months. RESULTS: Study of S. aureus, P. aeruginosa and BCC strains has shown, that 35 and 33.3% of cases of staphylococcus infection, 37 and 46% of pseudomonas infection in children and adults, respectively, 100% of BCC infections were determined by multi-resistant clones. Study of genotypically identical strains, isolated from a single patient at different stages, has shown a change in antibiotics sensitivity as a result of persistence. CONCLUSION: Persisent infection of lungs in patients with MV is determined: by exchanging clones with varying antibiotics sensitivity or prolonged circulation of a single clone with a high degree of phenotypical and genotypical variability, that determine alteration of seeding of sensitive and resistant strains from the same patient during monitoring. This confirms the necessity of study of antibiotics sensitivity of strains for prescription of antibacterial therapy.

[Burkholderia cepacia persistence in patients with mucoviscidosis].

AIM: Study features of persistence of Burkholderia cepacia in mucoviscidosis patients. MATERIALS AND METHODS: In the period from 2008 to 2009, 56 B. cepacia strains isolated from children with mucoviscidosis were obtained. 114 medical histories of children with mucoviscidosis from various age groups were analyzed. The developed algorithm for identification and typing including phenotype and molecular biology methods was used to identify B. cepacia bacteria. Strain genotyping was carried out by RAPD-PCR with random oligonucleotide primer as well as pulse-electrophoresis. RESULTS: Persistence of associations ofmicroogranisms in 59.4% of cases was established to be the feature of persistent infection in mucoviscidosis. The feature of persistence of B. cepacia strains in patients with diagnosis ofmuco-viscidosis mixed form, severe course is persistence in association with Pseudomonas aeruginosa. B. cepacia bacteria that can persist in mucoviscidosis patients are characterized by resistance to many antibiotics. A prolonged (up to 1 year and 5 months) persistence of B. cepacia strains isolated from 1 patient was proven by using microflora monitoring of lower respiratory tract. CONCLUSION: B. cepacia bacteria may colonize lower respiratory tract of mucoviscidosis patients, persist for a long time and be transmitted between patients.

[Microbial population of lower respiratory tract in children from different age groups with cystic fibrosis].

AIM: To study microflora of lower respiratory tract of children from different age groups with cystic fibrosis during follow-up for determination of its variability and possible sources of infectious complications. MATERIALS AND METHODS: One hundred forty-one medical histories of patients from different age groups with cystic fibrosis living in various regions of Russian Federation were analyzed. Eighty-four children with cystic fibrosis living in Moscow and Moscow region treated as outpatients and inpatients were prospectively followed. For identification and characterization of microorganisms, microbiological, molecular biological, and statistical methods were used. RESULTS: It was demonstrated that chronic pseudomonas, staphylococcal or mixed infection was already diagnosed in 25% of children aged 1-4 years, and identified in 80% of patients to the age of 18 years. In two-thirds of cases association of microorganisms was identified, and in hospitalized patients these associations were comprised by 3-5 microorganisms in 60% of cases. Aside from main agents in associations (Pseudomonas aeruginosa and Staphylococcus aureus), representatives of Gram-negative nonfermentative microorganisms (Burkholderia cepacia, Stenotrophomonas maltophilia, Acinetobacter baumanii) were often identified that possibly determined by tropism of these species to lung tissue. CONCLUSION: Chronic mixed infection is characteristic for patients with cystic fibrosis. Identification of possible mechanisms of lung infection in patients with cystic fibrosis will allow to develop evidence-based system of prevention of infectious complications in these patients.

[Role of "quorum sensing" regulatory system in formation of biofilms by Burkholderia cepacia and Pseudomonas aeruginosa].

AIM: To study influence of chemically synthesized lactones with different length of carbonic chain on formation of biofilms by Burkholderia cepacia and Pseudomonas aeruginosa. MATERIALS AND METHODS: Fifty-four strains of B. cepacia including reference strains and clinical isolates as well as etalon strain PA103 of P. aeruginosa were used. Lactones C4 [N-(3-hydroxybutanoyl-L-gomoserine lactone], C6 [N-(3-oxoohexanoyl)-L-gomoserine lactone], C8 [N-(3-oxooctanoyl)-L-gomoserine lactone], C0 [L-gomoserine lactone-HBr] were synthesized and purified by column chromatography. Formation of biofilms was studied by determination the ability of B. capacia strains to adhesion on the surface of 96-well polystyrene plate. RESULTS: Ability to biofilms formation was identified in 83.3% studied strains. It was shown that lactones C4+C8 and C6+C8 when added to cultivation medium improve growth of B. cepacia biofilm. Analysis of optical density (OD) values for P. aeruginosa biofilm revealed that lactones C4, C8, C4+C6, C4+C8, C6+C8 inhibit the formation of biofilm. The most prominent decrease of P. aeruginosa biofilm OD was observed during growth of culture in presence of C0. CONCLUSION: Obtained data point to different effects of lactones and their combinations on formation of biofilms by B. cepacia and P. aeruginosa. Suppression of biofilm formation by P. aeruginosa induced by lactone C0 confirms the need for development of new bacteriostatic drugs, which will be able to inhibit function of the "quorum sensing" regulatory system.

[Biofilm formation by strains of Burkholderia cepacia complex in dependence of their phenotypic and genotypic characteristics].

Biofilm formation was studied in 54 strains of Burkholderia cepacia complex isolated in 7 Moscow hospitals. 80% of strains (biofilm groups I and II) had the capacity to biofilm formation and only 16.7% of strains (group III) were not capable to biofilm formation. Molecular genetic methods allowed to identify one of the epidemic markers (CBL, IS hybrid sequence, Burkholderia Cepacia Epidemic Strain Marker - BCESM) in 46.7, 23.3, and 33.3% of strains from group I, II, and III respectively. Gene cepR from the Quorum Sensing system was identified in three biofilm groups in nearly equal frequency (92.3, 96.2 and 100% for group I, II, and III respectively), whereas cepl gene was found more often in group I (76.9%) and II (65.4%). Strains from all three groups had protease and lipase activity and 13.3% of group I strains had chitinolytic activity. B. cepacia strains from group I produced hemolysin in 33.3% of cases, from group II--in 26.6%, and from group III--in 11.1% of cases. The majority of Moscow hospital strains of B. cepacia complex were identified as B. cenocepacia (genomovar III, group A). RAPD-PCR method enabled to differentiate isolated strains into several genotypic variants. 13.3% of strains from group I were susceptible to imipenem/ciprofloxacin, as well as 33.3% of isolates from group II and 44.4% of isolates from group III.

[Antibiotic resistance and putative origin of Staphylococcus aureus and Klebsiella strains isolated from the children with intestinal dysbacteriosis].

The results of the statistical treatment of data on the analyses of 766 children, the residents of Moscow, for dysbacteriosis are presented; of these children, 34 were aged up to 1 month and 732, from 1 month to 1 year. This study revealed that in the fist year of life in children with dysbacteriosis the dominating bacterial species were S. aureus, bacteria of the genus Klebsiella and fungi of the genus Candida. From the intestine of children aged up to 1 month S. aureus and Klebsiella were isolated more often than from children aged up to 1 year. The results of the study of antibioticograms demonstrated that 21.6% of S. aureus strains and 74.4% of Klebsiella strains were multiresistant to antibiotics. Taking into account the fact that multiresistance to antibiotics was characteristic of hospital strains, the suggestion was made that the isolated strains were of hospital origin and such strains could colonize the intestine of children in maternity hospitals.

[Microbiological aspects of enteric dysbacteriosis in patients of different age groups, visiting outpatient clinics of Moscow].

The analysis of the intestinal microflora in 2,378 patients of different age revealed changes in the state of enteric microflora in all examined patients. In the maximum percent of cases a decrease in the amount of bifidobacteria was observed in children of up to 1 month old and in the amount of lactobacilli, in children aged 6 - 14 years. In patients of all age groups the representatives of such facultative microflora as Staphylococcus aureus or fungi of the genus Candida dominated. The highest proportion of isolated staphylococci was characteristic of children in the first year of life. In the highest percent of cases a decrease in the amount of Escherichia coli with typical properties was observed in persons over 65 years old. Other enterobacteria were most often isolated from adults aged 56 - 65, but the percentage of their isolation was 1.5 times lower than that of Candida. The conclusion was made that the treatment of patients with quantitative and qualitative disturbances of normal enteric microflora needed individual approach in each concrete case with due regard to the patient's age.

[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.

[Treatment of acute pain syndrome]. / Lechenie ostrogo bolevogo sindroma.

The article reviews modern concepts regarding stress response and its effects on various organs and systems of critical patients. Postoperative pain is regarded with emphasis on its pathophysiology and the prospects of using epidural anesthesia-analgesia for alleviating its injurious effects. Analgesia and sedation in patients with multiple injuries is discussed. The importance of adequate analgesia for alleviation of the injurious effects of nociceptive impulses in critical patients is emphasized. Analgesia should be regarded as one of the methods of intensive care of critical patients along with respiratory support, fluid replacement, and parenteral nutrition.

[Features of hormonal status in men during long-term coma]. / Osobennosti gormonal'nogo statusa pri dlitel'noi kome u muzhchin.

Serum hormonal profiles were studied in 48 patients during long (6-56 days) coma. Low T3 and T4 and an essential increase in the levels of hydrocortisone, prolactin, estradiol, and particularly progesterone (20-fold) in the presence of normal LH concentration and decreased level of FSH were observed on days 7-16. Changes in the concentrations of sex hormones, similar to the "false pregnancy" condition, were the most pronounced in the patients who later died.

[Use of semax at a follow-up of patients with posthypoxic encephalopathy]. / Primenenie semaksa v otdalennom periode u bol'nykh s postgipoksicheskoi patologiei mozga.

Effect of a neuropeptide semax is studied in 73 patients with various symptoms of posthypoxic encephalopathy in remote period. Fourteen of these patients developed a persistent vegetative state. The therapy was effective in patients with mnestic disorders; the drug effect was potentiated by an adaptogen oxiterm and by supraorbital electrophoresis. In some cases EEG showed episodes of paroxysmal activity after semax injection, and therefore, the first injection should be carried out with monitoring bioelectric activity of the brain.

[Dynamics of bioelectric activity of the brain in the acute period of stroke and its prognostic significance]. / Dinamika bioélektricheskoi aktivnosti mozga v ostrom periode insul'ta i ee prognosticheskaia znachimost'.

The electrical activity of the brain was assessed in various periods of acute ischemic stroke and its values correlated to the neurological symptoms. Seventy-six patients with acute ischemic stroke in the median cerebral artery basin were examined. A grave course of the disease and slowly progressing positive shifts in the neurologic symptoms were associated with the phenomenon of clinico-EEG-dissociation: an increase in the intensity of abnormally slow activity was wave-like and recorded both in the involved and intact hemispheres. These pathological shifts were maximally manifest at the site of projection of ischemic stroke on days 3, 7, and 14 of acute disorder of brain circulation, this permitting the use of EEG parameters in these days with prognostic purposes.

[Effect of the nude mutation on alpha-fetoprotein synthesis and hemopoiesis in the mouse liver in the postnatal period of development]. / Vliianie mutatsii nude na sintez al'fa-fetoproteina i gempoéz v pecheni myshei v postnatal'nom periode razvitiia.

The dynamics of population of alpha-fetoprotein (AFP)-containing cells in the liver and the level of AFP in the blood of C3H/HeJ+/+ and thymus-less mutant C3H/HeJnu/nu mice during postnatal development was studied by means of indirect immunofluorescence and radial immunodiffusion. The content of AFP-positive hepatocytes and AFP concentration in the blood serum of C3H/HeJnu/nu mice were shown to exceed markedly those in C3H/HeJ+/+ mice beginning from the age of 2 weeks. The histological analyses has revealed the foci of hemopoiesis in the liver of adult C3H/HeJnu/nu mice, unlike in the liver of normal mice. The neonatal thymectomy of C3H/HeJ+/+ mice did not influence the parameters under study. A possible relationship between the increased AFP level and the preservation of hemopoiesis in the liver of the mice homozygous by the mutation nude is discussed.

[Neurologic disorders in the late period following terminal states]. / Nevrologicheskie narusheniia v otdalennom periode posle terminal'nykh sostoianii.

The dynamics of the psychoneurological status f 70 patients who had experienced clinical death (17 patients), massive blood loss with prolonged hypotension (30 patients), and terminal eclampsia (23 patients) was studied. In 71% of the patients various late psychoneurological disorders were revealed. Distribution of these disorders according to the WHO classification is presented. It has been found that the psychoneurological disorders of this kind may either: a) gradually disappear, b) stabilize, or c) progress. In 42% of the patients new psychoneurological disorders appeared in the late period: such a deterioration became manifest 3 to 5 months after the terminal state sustained. The discovered incompleteness of the compensation processes points to a necessity of developing a rehabilitation program.

[Carcinogenicity of orthoaminoazotoluene for mouse intestines]. / O kantserogennosti ortoaminoazotoluola dlia kishechnika myshei.

Carcinogenic activity of orthoaminoazotoluene in relation to intestinal tissues of A/He mice was revealed. Intestinal tumours developed in 19 of 60 mice given this carcinogen; the tumours were localized in the cecum and represented adenomas and adenocarcinomas secreting mucus.