MORPHOLOGICAL CHARACTERISTICS OF SMALL INTESTINE MUCOSA IN DYSBIOSIS AND AFTER ITS CORRECTION BY PROBIOTICS AND ENTEROSORBENTS.

This study was aimed at investigating morphological and functional changes in the small intestine mucosal layer of mice with antibiotic-induced dysbiosis and following its correction with probiotics and enterosorbents. The study was carried out on BALB / c line white laboratory mice. Samples of the small intestine, liver, and spleen were taken to be processed for electron microscopy. To determine qualitative and quantitative composition of intestinal lumenal microbiome, animals' faeces were studied. Bacteria were isolated and identified by standard methods. This study has experimentally confirmed the ability of antibacterials to induce dysbiotic conditions in animals that are accompanied by significant shifts in the composition of normal microflora, manifested with cytodestructive disorders in the small intestinal epithelium. We have demonstrated the property of probiotics and, to a lesser extent, of sorbents to reduce the intensity and extension of cytodestructive disorders in the course of antibiotic-induced dysbiosis and to normalize the body immune responses that accompany the development of dysbiotic conditions.

Type of pH sensitive linker reveals different time-dependent intracellular localization, in vitro and in vivo efficiency in alpha-fetoprotein receptor targeted doxorubicin conjugate.

Despite the presence of a variety of modern anticancer drugs at the market, doxorubicin (Dox) is still widely used in antineoplastic therapy, although its administration causes severe side effects. To enhance specific activity of such molecules, various approaches have been exploited: targeted moieties like monoclonal antibodies, onco-specific proteins and peptides are utilized as specific vector molecules; environment sensitive linkers are exploited to facilitate transported drug release at the target point etc. Acid-labile linkers are frequently used in synthesis due to the ability to be cleaved inside specific cellular compartments with acidic environment, avoiding possible recycling mechanisms. Two types of conjugates containing different acid-labile linkers have been synthesized. In vitro efficiency of doxorubicin conjugates with recombinant receptor-binding domain of human alpha-fetoprotein (3dAFPpG) synthesized with use of cis-aconitic anhydride (CAA) and linker based on succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and 3-(2-pyridyldithio)propionic acid hydrazide (PDPH) was compared. The 3dAFPpG-SPDP-PDPH-Dox revealed a comparable with unmodified doxorubicin cytotoxic effect against the Dox sensitive MCF7 cell line and greater cytotoxicity against the anthracycline resistant MCF7Adr cells. Meanwhile the 3dAFPpG-CAA-Dox cytotoxic effect was significantly lower, although doxorubicin's pH-dependent release profiles and intracellular accumulation rates were similar. These differences in cytotoxic activity were arguably explained by the dissimilarities in intracellular doxorubicin localization, which may originate from thiol reductase activity in lysosomes and late endosomes.

Validation of RNA extraction procedures focused on micro RNA expression analysis.

The sampling procedure is a crucial step in every kind of experiment. This is also true in gene expression profiling experiments, where high quality and sufficient quantity of extracted RNA plays a significant role in the experimental outcome. We have compared five different RNA extraction protocols from peripheral blood/PBMCs with the aim to define the most suitable method for the miRNA expression profiling experiments. Convincing results in terms of highest quantity and quality were obtained by the TRIzol-chloroform extraction method. The total RNA obtained using this method contained the highest portion of good-quality miRNA molecules, which was also confirmed by gene-specific real-time PCR experiments.

[Antibacterial activity of aspen bark extracts against some pneumotropic microorganisms].

Extractive substances obtained from the bark of aspen (Populus tremula L.) with the use of petroleum ether (lipids I) and diethyl ether (lipids II) have exhibited high antibacterial activity with respect to Streptococcus pneumoniae and Haemophilus influenzae, causing 100% cell destruction. The minimum inhibiting concentration for S. pneumoniae has been found to be 0.5 - 50 mg/ml for lipids I and 0.0005 - 0.5 mg/ ml for lipids II, depending on the strain of bacteria. The antibacterial activity of rhytidome extracts is somewhat higher than that of phloem extracts. To suppress the growth of H. influenzae, more concentrated solutions of these extracts from 30 to 50 mg/ml are needed. Staphylococcus aureus was resistent in lipids. The action of temperature, mineral acids and alkali on lipids, as well as prolonged storage of strains in a refrigerator decreases the antibacterial activity of extracts under study.

[Etiological structure of respiratory infections in different variants of acute bronchitis]. / Etiologicheskaia struktura respiratornykh infektsii pri razlichnykh variantakh ostrogo bronkhita.

The microbiological and virological examination of 87 acute bronchitis patients (36 patients with the prolonged course and 31 patients with the relapsing course of the disease) was carried out. All forms of bronchitis were characterized by a high degree of infection with respiratory viruses and pneumococci. Haemophilus influenzae (type b) infection was registered rather rarely and only in combination with pneumococcal one. The highest characteristics of viro-bacterial associations were found in patients with acute bronchitis and with prolonged form of acute bronchitis, viral associations--with the prolonged and relapsing forms of the course of acute bronchitis in the presence of the bronchoobstructive syndrome.

[Bacterial infectious process in patients with chronic obliterating bronchiolitis]. / Bakterial'nyi infektsionnyi protsess u bol'nykh khronicheskim obliteriruiushchim bronkhiolitom.

Chronic obliterating bronchiolitis in children is characterized by the presence of chronic multifactor inflammatory infectious process with Streptococcus pneumoniae and Haemophilus influenzae being the leading causative agents. The inflammatory process is induced and maintained by S. pneumoniae and H. influenzae or by the association of these two pathogens, but the severity of the clinical course of obliterating bronchiolitis is determined by H. influenzae, which is confirmed by its high detection rate at the phase of exacerbation in patients with pronounced bronchial obstruction.

[Effect of halogen aerosol therapy on resistance parameters of the respiratory tract]. / Vliianie galoaérozol'noi terapii na zashchitnye svoistva respiratornogo trakta.

AIM: Assessment of the efficacy of dry high-dispersive aerosol of sodium chloride--the main acting factor of haloaerosol therapy--on defense system of the respiratory tract. MATERIAL AND METHODS: 188 patients with respiratory disease and at risk of pulmonary pathology received course of haloaerosol therapy. 49 matched patients were given placebo. The effect of the treatment was assessed by clinicoendoscopic picture, cytomorphological and bacteriological characteristics of the bronchoalveolar lavage, contamination activity of the microflora, activity of local humoral immunity in pharyngeal brush-biopsies and saliva, rheological indices of the sputum. RESULTS: Dry aerosol of sodium chloride demonstrated antiinflammatory activity in the respiratory tract, mucoregulating action. It enhances drainage of the bronchi, activates alveolar macrophages, improves biocenosis and local humoral immunity. CONCLUSION: Haloaerosol therapy has positive effect on the defense system, improves function of the respiratory tracts.

[Outcomes of the long-term treatment with a novel mucolytic drug pulmozin in mucoviscidosis patients]. / Opyt dlitel'nogo primeneniia novogo mukoliticheskogo preparata "pul'mozim" u bol'nykh mukovistsidozom.

AIM: To study clinical effectiveness and safety of a new mucolytic drug pulmozim (Switzerland) adjuvant to basic therapy in mucoviscidosis patients of different age in Russia. MATERIAL AND METHODS: 15 patients with mucoviscidosis aged 5-36 years, functional lung capacity and FEV-1 at least 40% received pulmozim for 30 days. The drug was given in a single daily dose 2.5 mg in inhalations. The study included three stages: initial 14 days--discontinuation of all mucolytic drugs, 30-day course of pulmozim, 14-day follow-up without pulmozim and other mucolytics. RESULTS: Pulmozim in the above regimen was effective in patients with mixed and moderate mucoviscidosis. The drug facilitates respiration, improves sputum rheology, normalizes general condition of the patient, is simple for use, has good organoleptic properties. CONCLUSION: Pulmozim is an effective mucolytic drug in combined treatment of mucoviscidosis.

[The use of halotherapy for the rehabilitation of patients with acute bronchitis and a protracted and recurrent course]. / Primenenie galoterapii dlia reabilitatsii bol'nykh ostrym bronkhitom s zatiazhnym i retsidiviruiushchim techeniem.

Halotherapy was used for rehabilitation in 25 patients with acute bronchitis of long-standing and recurrent types. The main therapeutic action was ensured by aerodispersed medium saturated with dry highly dispersed sodium chloride aerosol, the required mass concentration being maintained in the range of 1 to 5 mg/m3. Therapy efficacy was controlled through assessment of clinical, functional, immunological and microbiological findings. Metabolic activity values were taken into consideration as well. Positive dynamics of the function indices in the clinical picture resulted from elimination of pathogenic agents, control of slowly running inflammatory lesions and stimulation of some immune system factors. Favourable changes in metabolic activity were present: normalization of serotonin excretion, marked decrease of unbalance in lipid peroxidation-antioxidant system.

[Haemophilus influenzae biovars in patients with acute and chronic inflammatory lung diseases]. / Biovary Haemophilus influenzae u bol'nykh ostrymi i khronicheskimi vospalitel'nym zabolevaniiami legkikh.

In the study of the biological properties of 175 H. influenzae strains isolated from patients with acute and chronic inflammatory lung diseases (ILD) and from donors, a wide spread of biovars I, II and III (according to Kilian) was revealed; these biovars constituted 80% of the cultures under study. In donors, H. influenzae strains were characterized by a wide spectrum of biovars, but biovars I, II and VI constituted more than a half (64.2%) of the strains obtained in the course of these investigations. In acute ILD, only H. influenzae biovars I, II and III were isolated with the prevalence of biovar II (56.4%). In chronic ILD, all H. influenzae biovars were represented, but biovars II and III prevailed (58.7%). The four-fold difference in the occurrence of H. influenzae strains belonging to undetermined biovars was established in donors in comparison with ILD patients (46.7 +/- 9.8% and 12.0 +/- 2.5%; p less than 0.001).

[Antibiotic sensitivity of Haemophilus influenzae isolated from patients with inflammatory lung diseases]. / Chuvstvitel'nost' k antibiotikam Haemophilus influenzae, vydelennykh ot bol'nykh vospalitel'nymi zabolevaniiami legkikh.

In 1980-1986 the sensitivity of 2,045 H. influenzae strains isolated from the bronchial contents of patients with inflammatory lung diseases were studied. This study revealed that 60-80% of H. influenzae cultures circulating in Leningrad were sensitive to tetracyclin, chloramphenicol and erythromycin. During the period of observation the tendency towards the decrease of the number of highly sensitive H. influenzae cultures and the increase of the number of strains resistant to all antibiotic preparations was followed. Most of H. influenzae strains isolated in Leningrad were sensitive to penicillin, oleandomycin and ampicillin. In 1983 the appearance of H. influenzae strains, multiresistant to antibiotics, was noted. In 1986 these strains constituted 4.5% of all isolated cultures.

[Epidemiologic problems of pneumococcal infection]. / Nekotorye voprosy épidemiologii pnevmokokkovoi infektsii.

During the microbiological examination of 314 tentatively healthy donors, residents of Leningrad, Streptococcus pneumoniae was detected in the rhinopharynx of 94 donors (29.9%) in amounts ranging from a few individual microbial cells to 10(6) microbial cells with the mean geometric concentration equal to 3.58 lg. Antipneumococcal antibodies were detected in the blood of 147 donors (46.8%). The occurrence of antipneumococcal antibodies was approximately 2.5 times higher than the average level of carriership, but in most cases the dynamics of immunological reactions changed in the same direction. Considerable monthly and annual fluctuations in the occurrence of pneumococcal carriership and antipneumococcal antibodies, as well as their correlation with the epidemic rises of acute respiratory diseases, influenza and other acute respiratory viral diseases, were established. These data indicate that pneumococcal infection plays a definite role in the epidemic rises of acute respiratory diseases.

[Etiology of acute pneumonia occurring against a background of influenza and other acute respiratory viral infections]. / Etiologiia ostroi pnevmonii, protekaiushchei na fone grippa i drugikh ostrykh respiratornykh virusnykh infektsii.

The etiology of acute pneumonia (AP) was studied in 229 patients who had the disease simultaneously with influenza (106 patients), other viral and mycoplasmal infections (48 patients), and without concomitant acute viral infections (75 patients). The use of the quantitative microbiological method and the indirect immunofluorescence test with autostrains or Streptococcus pneumoniae strains of serotypes 2, 3, and 6, prevailing in Leningrad in patients with acute inflammatory diseases of the lungs, made it possible to find out the pneumococcal etiology of AP in 95% of patients irrespective of the presence of acute respiratory viral infections. The etiological role of opportunistic bacteria was revealed in 13 AP patients (5.7%); in 2 of them the causative agent of AP was Staphylococcus aureus and in 11, various species of Gram-negative enterobacteria. The latter were the cause of complications in 8 cases of pneumococcal pneumonia.

[Etiology of the infectious process in chronic pneumonia in children]. / Etiologiia infektsionnogo protsessa pri khronicheskoi pnevmonii u detei.

During the phase of exacerbation in chronic pneumonia in children Streptococcus pneumoniae and/or Haemophilus influenzae were isolated practically from all patients under examination. In the mild form of chronic pneumonia in children the high occurrence of H. influenzae (65.5-69%) was observed. During the whole acute phase of the disease, irrespective of the activity of the infectious process and the character of therapeutic measures, the degree of contamination of bronchial washings from the patients with these microorganisms was 5.95-6.6 lg/ml. In the bronchiectatic variant of chronic pneumonia the high activity and stability of the infectious processes induced by S. pneumoniae and H. influenzae were found to be linked with the considerable spread of these processes or with deeper morphological and functional changes in the bronchial stem and branches. During the phase of clinical remission in chronic pneumonia in 36.8-58.8% of children the release of S. pneumoniae and H. influenzae was observed. The geometric mean of their concentration varied within 3-5.3 lg/ml.

[Antibiotic sensitivity of Streptococcus pneumoniae strains isolated from the bronchial contents of patients with inflammatory lung diseases]. / Chuvstvitel'nost' k antibiotikam shtammov Streptococcus pneumoniae, vydelennykh iz bronkhial'nogo soderzhimogo bol'nykh s vospalitel'nymi zabolevaniiami legkikh.

The study of the sensitivity of 2681 pneumococcal strains to antibiotics, carried out in 1976-1982, demonstrated that most of these strains circulating in Leningrad were sensitive to penicillin, levomycetin, erythromycin and oleandomycin. Penicillin-resistant pneumococcal strains were found to appear in 1980 and later showed a definite increase in number, reaching 4.4% of all strains in 1982. Simultaneously the appearance of levomycetin- and erythromycin-resistant strains and the increase of their relative significance were established. Most of the pneumococcal strains isolated in Leningrad were not sensitive to monomycin, streptomycin and tetracyclin.

[Pneumococcus serotypes in acute and chronic inflammatory lung disease (1978-1980)]. / Serotipy pneuvmokokka pri ostrykh i khronicheskikh vospalitel'nykh zabolevanii legkikh (1978-1980).

The comparison of the antigenic structure of the polysaccharide capsule in 342 pneumococcal strains isolated in Leningrad from the bronchial contents of patients with acute and chronic inflammatory pulmonary diseases indicated that during 1978-1980 a change in the composition of the prevailing groups of serotypes occurred every year. The comparison of the groups of prevailing pneumococcal serotypes isolated from children and adults has revealed no statistically significant differences in the specific prominence of different types, with the exception of serotype 15. Serotypes 6, 7 and 2 have been found to occur most frequently in acute pneumonia, and serotypes 23, 3, 9, 6, 15, 29 and 34 in chronic inflammatory pulmonary diseases. The preliminary data on the differences in the composition of the prevailing pneumococcal serotypes causing acute pneumonia in Leningrad and Kaunas have been obtained.

[Microbiological methods of diagnosing pneumococcal infection in acute and chronic inflammatory lung disease]. / Mikrobiologicheskie metody diagnostiki pnevmokokkovoi infektsii pri ostrykh i khronicheskikh vospalitel'nykh zabolevaniiakh legkikh.

The simultaneous microbiological study of sputum and bronchial washings from 440 patients with acute and chronic inflammatory pulmonary diseases has revealed that pneumococci can be isolated most frequently from both these materials or sputum alone. The isolation of pneumococci by the inoculation of the pathological materials or sputum alone. The isolation of pneumococci by the inoculation of the pathological material into solid culture media is practically as effective as the inoculation of white mice with this material, but the former method accelerates the isolation of Str. pneumoniae and allows one to determine the content of these organisms in the initial material. The appearance and the size of pneumococcal colonies depend on the composition of the culture medium, the biological properties and the amount of Str. pneumoniae and other microorganisms in the material under study. The identification of the S-forms of pneumococci is based on their cultural and morphological properties and carried out by one test or by a complex of tests. 4 criteria allowing to determine the etiological importance of pneumococci isolated from patients are proposed.