Assessment of cancer stem cell marker expression in primary head and neck squamous cell carcinoma shows prognostic value for aldehyde dehydrogenase (ALDH1A1).

Cancer stem cells (CSCs) play a key role in carcinogenesis and progression of head and neck squamous cell carcinomas (HNSCC). The most common markers indicating for CSCs are: CD44, CD24, CD133, ALDH1A1. Our objective was to evaluate the prognostic potential of CSC markers in HNSCC. The study included 49 patients treated for primary HNSCC, 11 patients with upper respiratory tract epithelial dysplasia and 12 subjects with the normal pharyngeal mucosa as a control group. The frequency and expression levels of the four CSC markers were assessed by immunohistochemistry. Univariate and multivariate analyses were used to correlate CSC expression levels with tumor stage, lymph node metastases or overall survival (OS). CD44, CD24, CD133, ALDH1A1 were widely expressed in tumors, whereas CD44 was found to be higher in cancer tissue (P = 0.001). ALDH1A1 expression levels were found to be significantly higher in T3-T4 tumors vs. T1-T2 tumors (P = 0.05). Lymph node metastases had significantly higher expression levels of CD24 (P = 0.01) and CD133 (P < 0.05) than primary tumors. Multifactorial analysis revealed that overall survival (OS) for patients with ALDH1A1 negative tumors was 5.25 times higher than for patients with ALDH1A1 positive (ALDH1A1+) tumors (P = 0.01). On univariate and multivariate analysis, only ALDH1A1 positivity had a significant effect on OS of HNSCC patients (HR = 2.47 for P = 0.02). Immunohistochemistry-based assessments of CSC marker expression in HNSCC has significant predictive implications for patients with HNSCC. The frequency of CSCs in the tumor, specifically of ALDH1A1+ cells correlated with five-year OS in these patients.

Systems pharmacology-based study of Tanreqing injection in airway mucus hypersecretion.

ETHNOPHARMACOLOGICAL RELEVANCE: Mucus hypersecretion (MH) is recognized as a key pathophysiological and clinical feature of many airway inflammatory diseases. MUC5AC is a major component of airway mucus. Tanreqing injection (TRQ) is a widely used herbal formula for the treatment of respiratory inflammations for years in China. However, a holistic network pharmacology approach to understanding its therapeutic mechanisms against MH has not been pursued. AIM OF THE STUDY: This study aimed to explore the systems-level potential active compounds and therapeutic mechanisms of TRQ in the treatment of MH. MATERIALS AND METHODS: We established systems pharmacology-based strategies comprising compound screenings, target predictions, and pathway identifications to speculate the potential active compounds and therapeutic targets of TRQ. We also applied compound-target and target-disease network analyses to evaluate the possible action mechanisms of TRQ. Then, lipopolysaccharide (LPS)-induced Sprague-Dawley (SD) rat model was constructed to assess the effect of TRQ in the treatment of MH and to validate the possible molecular mechanisms as predicted in systems pharmacology approach. RESULTS: The comprehensive compound collection successfully generated 55 compound candidates from TRQ. Among them, 11 compounds with high relevance to the potential targets were defined as representative and potential active ingredients in TRQ formula. Target identification revealed 172 potential targets, including pro-inflammatory cytokines of tumor necrosis factor α (TNF-α), interleukin (IL)-6, and IL-8. Pathway analyses uncovered the possible action of TRQ in the regulation of IL-17 signaling pathway and its downstream protein MUC5AC. Then in vivo experiment indicated that TRQ could significantly inhibit LPS stimulated MUC5AC over-production as well as the expression of TNF-α, IL-6, IL-8, and IL-17A, in both protein and mRNA levels. CONCLUSIONS: Based on the systems pharmacology method and in vivo experiment, our work provided a general knowledge on the potential active compounds and possible therapeutic targets of TRQ formula in its anti-MH process. This work might suggest directions for further research on TRQ and provide more insight into better understanding the chemical and pharmacological mechanisms of complex herbal prescriptions in a network perspective.

A novel rat model for tracheal mucosal damage assessment of following long term intubation.

OBJECTIVE: Tracheal mucosal damage is a well-known complication of endo-tracheal intubation and animal models are essential for studying the underlying cellular injury cascade. The novel rat model described here is based on retrograde intubation via tracheotomy and suture fixation of the tube. It aims to simulate the common clinical scenario of tube-related airway damage due to long term intubation. STUDY DESIGN: Prospective randomized control pilot study. METHODS: Male Sprague-Dawley were randomly assigned into two groups: control (no intubation, n = 10), one week of intubation (n = 13). The animals were then euthanized and the trachea was sent for histological analysis. Epithelial damage, mucosal thickness, mucosal gland hypertrophy and fibrosis were reviewed. RESULTS: Intubation procedure survival rate was 84.6% (11/13) and 100% in the control (10/10). The damaged ciliary mechanism was a common finding in the intubated group compared to the preserved normal ciliary architecture in almost all control rats. Average tracheal mucosal thickness was 119.0 ±â€¯21.8 µm for the control group and 254.6 ±â€¯22.8 µm for the intubated group, (p < 0.001). The ciliary damage score was 1.00 ±â€¯0.02 in the intubated group, and 0 ±â€¯0.02 in the control group. (p < 0.001). The (objective) average total tracheal mucosal gland area was 19,530 ±â€¯24,606 in the intubated group and 10,031 ±â€¯23,461 in the control group (p < 0,05). Collagen deposition seems higher in the intubated trachea compared to the control. CONCLUSIONS: We describe a novel rat-based animal model for simulating tracheal mucosal damage following long term intubation. This animal model is easy to carry out, reproducible and involves containable animal mortality rates. LEVEL OF EVIDENCE: I.

Experimental animal model for assessment of tracheal epithelium regeneration.

OBJECTIVES/HYPOTHESIS: To develop an experimental model in rabbits for assessment of tracheal epithelium regeneration through application of either natural or artificial polymer scaffolds. STUDY DESIGN: First, we identified the size of full-thickness mucosal defect, which does not allow self-healing (a "critical defect"), thus representing an adequate experimental model for regenerative therapy of tracheal epithelium damage. Then, two methods of polymer scaffold fixation at the site of the epithelium defect were compared: suturing and fixation with a stent. This was done through: 1) formation of a full-thickness anterolateral mucosal defect by tracheal mucosa excision; and 2) fixation of the scaffold at the site of the tracheal epithelium defect using sutures (through a tracheal wall "window") or a vascular stent (through a small tracheal incision). RESULTS: The dimension of a critical anterolateral mucosal defect of the trachea for rabbits was found to be 1.5 cm in length and more than 50% of the tracheal circumference. Fixation of the scaffold with a stent proved to be more efficient due to a uniform distribution of the pressure over the entire surface of the scaffold, whereas the suturing of the scaffold provided unsatisfactory results. In addition, fixation of the scaffold by suturing required formation of a large "window" in the tracheal wall. Thus, using the stent appeared to be technically less complicated and much less traumatic as compared to suturing. CONCLUSION: We present an experimental in vivo animal model of tracheal epithelium injury and recovery. It can be effectively used with certain further modifications as a basis for routine testing of bioengineered constructs. LEVEL OF EVIDENCE: NA Laryngoscope, 129:E213-E219, 2019.

A 104-week pulmonary toxicity assessment of long and short single-wall carbon nanotubes after a single intratracheal instillation in rats.

We compared long-term pulmonary toxicities after a single intratracheal instillation of two types of dispersed single-wall carbon nanotubes (SWCNTs), namely, those with relatively long or short linear shapes with average lengths of 8.6 and 0.55 µm, respectively. Both types of SWCNTs were instilled intratracheally in male F344 rats at 0.2 or 1.0 mg/kg (long SWCNTs) or 1.0 mg/kg (short SWCNTs). Pulmonary responses were characterized at 26, 52 and 104 weeks after a single instillation. Inflammatory changes, test substance deposition, test substance engulfment by macrophages, and alveolar wall fibrosis were observed in the lungs of almost all test rats at 52 and 104 weeks after short nanotube instillation. The incidences of these changes were much lower in the long nanotube-treated groups. In almost all rats of the long nanotube-treated groups, fibrosis and epithelium loss in the terminal bronchiole with test substance deposition were observed. These bronchiolar changes were not observed after administering short nanotubes. Both bronchiolo-alveolar adenoma and carcinoma were found in the negative-control group, the high-dose long-nanotube group, and the short-nanotube group at 104 weeks post-instillation, although the incidences were not statistically different. The genotoxicity of the SWCNTs was also evaluated by performing in vivo comet assays with lung cells obtained 26 weeks post-instillation. No significant changes in the percent tail deoxyribonucleic acid were found in any group. These findings suggested that most long SWCNTs were deposited at the terminal bronchioles and that a considerable amount of short SWCNTs reached the alveolus, resulting in chronic inflammatory responses, but no genotoxicity in the lungs.

[Theoretical basis and clinical benefits of dry salt inhalation therapy]. / A szárazsó-belégzéssel történo kezelés elméleti alapjai és gyakorlati haszna.

Dry salt inhalation (halotherapy) reproduces the microclimate of salt caves, with beneficial effect on health. Sodium chloride crystals are disrupted into very small particles (with a diameter less than 3 µm), and this powder is artificially exhaled into the air of a comfortable room (its temperature is between 20-22 °C, and the relative humidity is low). The end-concentration of the salt in the air of the room will be between 10-30 mg/m(3). The sick (or healthy) persons spend 30-60 minutes in this room, usually 10-20 times. Due to the greater osmotic pressure the inhaled salt diminishes the oedema of the bronchial mucosa, decreases its inflammation, dissolves the mucus, and makes expectoration easier and faster (expectoration of air pollution and allergens will be faster, too). It inhibits the growth of bacteria and, in some case, kills them. Phagocyte activity is also increased. It has beneficial effect on the well being of the patients, and a relaxation effect on the central nervous system. It can prevent, or at least decrease the frequency of the respiratory tract inflammations. It produces better lung function parameters, diminishes bronchial hyperreactivity, which is the sign of decreasing inflammation. Its beneficial effect is true not only in inflammation of the lower respiratory tract, but also in acute or chronic upper airways inflammations. According to the international literature it has beneficial effect for some chronic dermatological disease, too, such as psoriasis, pyoderma and atopic dermatitis. This treatment (called as Indisó) is available under medical control in Hungary, too.

Impact Assessment of Cigarette Smoke Exposure on Organotypic Bronchial Epithelial Tissue Cultures: A Comparison of Mono-Culture and Coculture Model Containing Fibroblasts.

Organotypic 3D cultures of epithelial cells are grown at the air-liquid interface (ALI) and resemble the in vivo counterparts. Although the complexity of in vivo cellular responses could be better manifested in coculture models in which additional cell types such as fibroblasts were incorporated, the presence of another cell type could mask the response of the other. This study reports the impact of whole cigarette smoke (CS) exposure on organotypic mono- and coculture models to evaluate the relevancy of organotypic models for toxicological assessment of aerosols. Two organotypic bronchial models were directly exposed to low and high concentrations of CS of the reference research cigarette 3R4F: monoculture of bronchial epithelial cells without fibroblasts (BR) and coculture with fibroblasts (BRF) models. Adenylate kinase (AK)-based cytotoxicity, cytochrome P450 (CYP) 1A1/1B1 activity, tissue histology, and concentrations of secreted mediators into the basolateral media, as well as transcriptomes were evaluated following the CS exposure. The results demonstrated similar impact of CS on the AK-based cytotoxicity, CYP1A1/1B1 activity, and tissue histology in both models. However, a greater number of secreted mediators was identified in the basolateral media of the monoculture than in the coculture models. Furthermore, annotation analysis and network-based systems biology analysis of the transcriptomic profiles indicated a more prominent cellular stress and tissue damage following CS in the monoculture epithelium model without fibroblasts. Finally, our results indicated that an in vivo smoking-induced xenobiotic metabolism response of bronchial epithelial cells was better reflected from the in vitro CS-exposed coculture model.

Assessment of the respiratory sensitization potential of proteins using an enhanced mouse intranasal test (MINT).

There remains a need for a simple and predictive animal model to identify potential respiratory sensitizers. The mouse intranasal test (MINT) was developed to assess the relative allergic potential of detergent enzymes, however, the experimental endpoints were limited to evaluation of antibody levels. The present study was designed to evaluate additional endpoints (serum and allergic antibody levels, pulmonary inflammation and airway hyperresponsiveness (AHR)) to determine their value in improving the predictive accuracy of the MINT. BDF1 mice were intranasally instilled on days 1, 3, 10, 17 and 24 with subtilisin, ovalbumin, betalactoglobulin, mouse serum albumin or keyhole limpet hemocyanin; challenged with aerosolized methacholine or the sensitizing protein on day 29 to assess AHR, and sacrificed on day 29 or 30. Under the conditions of this study, evaluation of AHR did not improve the predictive power of this experimental model. Allergic antibody responses and IgG isotype characterization proved to be the most sensitive and reliable indicators of the protein allergenic potential with BAL responses providing additional insight. These data highlight that the evaluation of the respiratory sensitization potential of proteins can be best informed when multiple parameters are evaluated and that further improvements and refinements of the assay are necessary.

Mucosal changes in laryngopharyngeal reflux--prevalence, sensitivity, specificity and assessment.

OBJECTIVES/HYPOTHESIS: A literature review regarding the use of laryngopharyngeal mucosal signs in diagnosing laryngopharyngeal reflux (LPR). STUDY DESIGN: Literature review. METHODS: A search of MEDLINE in February 2012 using the terms laryngopharyngeal reflux, laryngitis, mucosa, appearances, and signs (English language only). RESULTS: One or more laryngopharyngeal mucosal signs associated with LPR were identified in 64% to 93% of healthy volunteers (3% >5 signs) and in 17% to 85% of gastroesophageal reflux disease sufferers (Reflux Finding Score [RFS] >7 in 24%). Reinke's edema, pseudosulcus, ventricular obliteration, vocal cord nodules, and granulomas have in some, but not all studies, been shown to be more prevalent in those with pH-proven pharyngeal reflux. Pseudosulcus, interarytenoid thickening, and Reinke's edema were more prevalent in those symptomatic of LPR than those not. The use of multiple mucosal signs may improve detection of reflux sufferers from asymptomatic controls. The RFS has a sensitivity and specificity of 87.8% and 37.5%, respectively, for picking up pH-proven pharyngeal reflux individuals. Inter- and intrarater reliability for identifying signs is fair to good in most studies. CONCLUSIONS: The limited evidence for each mucosal finding should be considered in making the diagnosis of LPR. Further quality research in to mucosal findings in LPR is needed.

Relationship between immunohistochemical assessment of bronchial mucosa microvascularization and clinical stage in asthma.

Although hardly ever used in current practice, fibrobronchoscopy may provide interesting histopathological-clinical correlations in patients diagnosed with different stages of evolutive asthma. The aim of the study was to evaluate the correlation between semi-quantitative microvascularization features and the asthma severity assessed according to the GINA classification 2006. Our study group consisted in 21 patients diagnosed with asthma of different stages of severity and two-control patients investigated by fibrobronchoscopy with associated biopsy. The tissue fragments underwent standard processing procedures for the immunohistochemical exam, using CD34 as microvascularization marker. The semi-quantitative analysis was based on the "hot spot" method and on a score system that corresponds to the microvessels density. The statistical analysis of the correspondence between CD34 score and clinical parameters was performed using the SPSS 17 software, applying non-parametric correlation tests. The CD34 evaluation showed an increase in blood vessels count in all asthmatic patients in comparison to the control group and a close correlation with the asthma severity, reflected by the FEV1 values. The statistical analysis showed an inverse correlation between FEV1 [%] values and CD34 expression (r=-0.93, p<<0.01). Our data concur to other research reports, supporting the hypothesis that angiogenesis initially facilitates the edema development and later on appears to be involved in the bronchial wall thickening, as a component of the chronic inflammatory response, with concomitant distensibility reduction. The bronchial mucosa microvascularization evaluation opens new perspectives for advanced therapies, with beneficial effects for asthmatic patients' life quality.

[Health status assessment of the population living in vicinity of carrier rocket fall areas].

One hundred and eighteen subjects living at various distances from the carrier rocket fall places underwent cytomorphological studies of the upper airways (rhinocytogram) and buccal epithelium. The findings indicate that the dwellers of the settlements of Ulytausky District have chronic hypertrophic rhinitis since the nasal mucosa and the buccal epithelium are the first and most important biological barrier on the way of adverse and technogenic factors to come into the organism. Technogenic pollutants have cumulative activities, causing granulated mast cells to accumulate; then they actively secrete biologically active substances, by impairing the epithelial responsiveness in both the upper airways and the buccal epithelium.

[Computer method for the assessment of epithelial metaplasia in human bronchial mucous membrane].

Using Bio Vision computer program, the successive stages of metaplastic changes development were traced in the pseudostratified columnar ciliated epithelium of bronchial mucous membrane obtained from 65 patients with bronchial asthma of various degrees of severity. Biopsy material analysis demonstrated the changes of cell nuclear and cytoplasmic density, as well as of basement membrane.

Critical assessment of an in vitro bovine respiratory organ culture system: a model of bovine herpesvirus-1 infection.

A bovine in vitro organ culture (BIVOC) system was evaluated as a model to study host and pathogen events during the course of bovine herpesvirus-1 infection. Upper respiratory tract epithelium, from slaughtered animals, was cultured in an air-liquid interface system and integrity, viability, and TNF-alpha gene expression of tissue explants were monitored over 72h in the presence or absence of infection by bovine herpesvirus type 1 (BHV-1). Uninfected explants maintained viability and integrity over the 72h time course although histological signs of degeneration were first visible from 24h of culture. Explants were productively infected with BHV-1 and typical, dose dependent, cytopathic changes were observed in response to infection. Regulation of TNF-alpha gene expression in uninfected explants varied over time and was region-specific but there was significant down-regulation of TNF-alpha gene expression at 2h post-infection when compared to uninfected controls at the same time point. Taking caveats into consideration the BIVOC system shows promise as a tool for analysis of immediate or early events in host-pathogen interaction.

[Disturbance of the microenvironment of upper respiratory tract mucosa as an indicator of early changes in the health status under combined effect of toxic substances].

Results of quantitative and qualitative assessment of mucosal microbiocenoses in the upper respiratory tracts are presented along with cytomorphological characteristics of upper airway mucosa. SIgA and lysozyme concentrations were measured in the saliva of firemen. Their exposure to occupational hazards during periods of professional activity was shown to lead to a decrease in non-specific anti-infectious resistance to the effect of unfavourable factors.

Appropriate interslice gap for screening coronal paranasal sinus tomography for mucosal thickening.

The objective of this study was to establish the appropriate interslice gap for screening coronal paranasal sinus tomography to identify sinus mucosal thickening. We reviewed 100 coronal paranasal sinus tomographic scans (interslice gap, 2 mm) that had been performed at our institution between January 2004 and November 2004 to evaluate rhinosinusitis. Digital photographs of all slices from each tomographic scan were taken. The intervening slices were eliminated to form six different sets of interslice gaps of 4, 6, 8, 10, 16, and 20 mm. The remaining slices for each set were moved to corresponding folders created on a computer to catalog each interslice gap. The same specialist evaluated each folder of interslice gap. The paranasal sinuses, the ethmoid infundibulum, and the frontal recess were evaluated for mucosal thickening. The sensitivity, specificity, and accuracy of each interslice gap in detecting mucosal thickening were calculated by accepting the results of 2-mm-thick slices as the gold standard. The interslice gap of 2 mm was compared with that of other interslice gaps using the chi-square test for dependent groups (the McNemar test). The value of 20 mm interslice gap in detecting sinus mucosal thickening was found to be significantly low when compared with the interslice gap of 2 mm (P = 0.022). Using coronal paranasal sinus tomography, an interslice gap up to 16 mm may be used to detect sinus mucosal thickening.

In vitro models for the assessment of inflammatory and immuno-modulatory effects of the volatile organic compound chlorobenzene.

An in vitro cell culture system based on an air/liquid culture technique was developed which allows a direct exposure of cells to volatile chemicals without medium coverage. For the establishment of the experimental system, chlorobenzene was used as a model compound. Chlorobenzene is a volatile organic compound which is mainly used as a solvent. Beside other adverse health effects, chlorobenzene exposure has been shown to be associated with respiratory tract irritations, Th2 differentiation, and allergic sensitizations. Human peripheral blood mononuclear cells (PBMC) and lung epithelial cells (A549) were exposed to chlorobenzene via gas phase for 20 h. Additionally, PBMC were incubated with culture supernatants from exposed lung epithelial cells. High chlorobenzene concentrations (100 g/m(3)) induced IL-8 production in A549 cells, whereby lower concentrations (10 microg/m(3)-1 g/m(3)) stimulated the secretion of the monocyte chemoattractant protein-1 (MCP-1). A direct effect of chlorobenzene on the cytokine secretion of PBMC was not found. However, if PBMC were incubated with culture supernatants of exposed lung cells, an enhanced production of the Th2 cytokine IL-13 was observed. This induction was prevented in the presence of an anti-MCP-1 antibody. Our data suggest that chlorobenzene induces the production of inflammatory mediators in lung cells. The primary chlorobenzene caused release of MCP-1 in lung epithelial cells may secondarily result in a Th2 differentiation in T lymphocytes. These findings may contribute to the understanding of how chlorobenzene mediates the development of inflammatory reactions in the airways and contributes to the development of an allergic reactivity.

Comparative assessment of a metabolically attenuated Mycoplasma gallisepticum mutant as a live vaccine for the prevention of avian respiratory mycoplasmosis.

In a previous study, signature sequence mutagenesis (SSM) was used to identify a mutant with a disruption of the gene encoding the metabolic factor, dihydrolipoamide dehydrogenase, and that mutant was designated Mg 7. The current study assessed the safety, immunogenicity and efficacy of Mg 7 in comparison to two commercially available vaccines (ts-11 and F) as well as a laboratory vaccine strain, GT5. Intratracheal vaccination of chickens with all four attenuated mutants induced varying levels of protection against intratracheal challenge with virulent Mycoplasma gallisepticum strain R(low). Mg 7 vaccinated chickens rapidly cleared the challenge strain, had lower histopathologic tracheal lesion scores when compared to unvaccinated chickens, and mounted a strong humoral anti-M. gallisepticum-specific IgG response. The IgG levels increased 2- to 3-fold upon R(low) challenge. Mg 7 induced a greater level of protection against intratracheal R(low) challenge than that observed with the other three attenuated strains, as evidenced by a lower recovery of R(low) from tracheas and lower histopathologic lesion scores in tracheas and air sacs. Based on these findings, Mg 7 appears to have good potential as a safe and effective vaccine for the prevention of avian mycoplasmosis.

Exposure of bronchial epithelial cells to whole cigarette smoke: assessment of cellular responses.

Cigarette smoke is composed of approximately 5% particulate phase and 95% vapour phase by weight. However, routine in vitro toxicological testing of smoke normally only measures the activity of the particulate phase. This study describes a new system for exposing cells at an air-liquid interface to serial dilutions of gaseous smoke. Confluent monolayers of NCI-H292 human lung epithelial cells on semipermeable membranes were placed in a purpose-designed Perspex chamber at an air-liquid interface. The cells were exposed to dilute whole mainstream cigarette smoke for 30 minutes, followed by a 20-hour recovery period. Firstly, high and low delivery cigarettes were compared, and cytotoxicity was determined by using the neutral red uptake assay. Clear differential cytotoxic responses were observed with the two cigarette types, which correlated positively with the concentrations of components in smoke, and particularly compounds in the vapour phase, such as aldehydes. Secondly, low doses of smoke were found to up-regulate mRNA levels of the secreted mucin, MUC5AC, and to stimulate the production of interleukin (IL)-6, IL-8 and matrix-metalloprotease-1, but had no effect on growth-related oncogene alpha. This system will facilitate further investigations into the toxicological mechanisms of cigarette smoke components, and may be useful for studying other gaseous mixtures or aerosols.

Assessment of CFTR localisation in native airway epithelial cells obtained by nasal brushing.

Reliable methods for determining the localisation of mutant CFTR protein in native cells from CF individuals are necessary to allow the degree of mislocalisation of any genotype to be defined and to assess the effect of therapeutic agents on CFTR trafficking. Here, we present procedures for obtaining ciliated epithelial cells from CF patients by nasal brushing and a description of protocols for immunolocalisation of CFTR. The protocols are a consensus, following comparison of some aspects of methods currently used in the authors' laboratories.

[Clinicomorphological assessment of budesonide efficiency in patients with bronchial asthma]. / Kliniko-morfologicheskaia otsenka éffektivnosti budesonida u bol'nykh bronkhial'noi astmoi.

AIM: to study clinicomorphological efficacy of inhalation glucocorticosteroid budesonide (benacort, Pulmomed, Russia) in bronchial asthma. MATERIAL AND METHODS: Twenty patients with bronchial asthma were treated with budesonide. RESULTS: A response to budesonide was manifest to the end of treatment week 1. Budesonide reduced frequency of acute asthma episodes and the need in inhalations of short-acting beta 2-agonists. The peak expiratory velocity (PEV) rose by 12% in three months, variability of PEV lowered by 13%, in 6 months by 21%, in 12 months by 31% compared to pretreatment values. In 12 months hypersecretion and thickness of basal membrane decreased. Three-month treatment also reduced eosinophil and lymphocyte epithelial count and cell density of stromal infiltrate in bronchial mucosa. In 12 months cell density of stromal infiltrate diminished. CONCLUSION: Bronchial asthma treatment with budesonide for 12 months reduces 24 hour rate of acute asthma episodes, the need in the disease exacerbations, improves functional indices of respiration but morphological composition of bronchial mucosa does not normalize completely this showing the necessity of longer budesonide administration.