High prevalence of W1282x mutation in cystic fibrosis patients from Karachay-Cherkessia.

Cystic fibrosis (CF; OMIM #219700) is a common autosomal recessive disease. The spectrum and frequency of CFTR mutations vary significantly in different populations and ethnic groups. A genetic epidemiological study was conducted in the indigenous ethnic group of people known as the Karachais. They live in the Republic of Karachay-Cherkessia, which lies in the northwest of Russia's North Caucasus region. Karachai's are Turkic-speaking and consist of 194 thousand people (approximately 40% of the population of the Republic). Molecular genetic analysis was performed in 10 unrelated Karachai families with CF patients from three districts in the Republic. A high frequency of W1282X mutation was found (18 of 20 mutant alleles): eight patients were homozygous for the W1282X mutation, and two were compound heterozygous (the second alleles were R1066C and R709X). Analysis for 13 common CF mutations in the sample of 142 healthy Karachais identified two 1677delTA and two W1282X mutation carriers. Thus, the most common CFTR mutation, F508del, was not detected among the CF patients or in healthy Karachais. The most frequent mutation among Karachai patients is W1282X (90%). Its frequency in healthy Karachais is approximately 0.007. Haplotype analysis using the CFTR intragene DNA markers IVS1CA, IVS6aGATT, IVS8CA and IVS17bCA showed that the origins of the W1282X mutation in Karachay-Cherkessia and the Eastern European part of Russia are different.

Tumor necrosis factor-alpha/interleukin-10 balance in normal and cystic fibrosis children.

BACKGROUND: The balance between tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) is important for immune homeostasis maintenance. Exuberant production of TNF-alpha contributes to overwhelming inflammatory response and tissue damage. But, commonly, increase in TNF-alpha is counterbalanced by simultaneous synthesis of an anti-inflammatory cytokine IL-10, which suppresses production of many activating and regulatory mediators. AIMS: In the present study, the relationships between TNF-alpha and IL-10 in the plasma of healthy school-children and cystic fibrosis (CF) patients have been investigated. METHODS: Blood samples were obtained from 12 CF patients with chronic pulmonary disease and 18 healthy schoolchildren vaccinated with live attenuated rubella vaccine. IL-10 and TNF-alpha were determined in the plasma samples using commercially available enzyme-linked immunosorbent assay kits. RESULTS: Before vaccination, most healthy children (13 of 18) demonstrated superiority of pro-inflammatory TNF-alpha over anti-inflammatory IL-10 (TNF-alpha/IL-10 > 1). In these subjects, a significant positive linear association between the cytokine values has been found. Vaccine challenge resulted in a marked reduction of TNF-alpha/IL-10 ratios. In addition, a disappearance of correlation between the cytokine values was observed. Such disturbance was related to exuberant elevation of the IL-10 levels after inoculation. On the contrary, in CF individuals, plasma cytokine values remained in strong linear association independently of TNF-alpha or IL-10 predominance. No spikes in the plasma levels of IL-10 in CF patients during a 6-month observation period have been revealed. CONCLUSIONS: There were no fundamental differences between CF and healthy children in the regulation of TNF-alpha and IL-10 secretion. Thus, immune quiescence seemed to be associated with the predominance of TNF-alpha, whereas immune disturbance was characterized by IL-10 superiority. The only abnormality that was found in CF patients consisted of their inability to produce unlimitedly IL-10 in response to antigen stimuli.

[Antibiotic therapy of cystic fibrosis in children]. / Antibiotikoterapiia pri muskovistsidoze u detei.

It is postulated that P. aeruginosa in monoculture or in association with Staphylococcus aureus keeps its leading position in chronic bacterial inflammatory broncho-pulmonary processes in children with cystic fibrosis. Antibiotic resistant strains of Burkholderia cepacia, Stenotrophomonas maltophila, Alcaligenes xylosoxidans were revealed (7.1% of the strains). P. aeruginosa strains were susceptible to aminoglycosides, ciprofloxacin, and polymixin B. Susceptibility of smooth and mucoid forms of P. aeruginosa to ceftazidime stayed at the level of 49.6-57.1%. Such microbial associations as P. aeruginosa sm. + S. aureus, P. aeruginosa sm. + P. aeruginosa muc. + S. aureus were mainly susceptible to ciprofloxacin, aminoglycosides and resistant to ceftasidime. Meropenem, cefepim and ciprofloxacin are highly effective antibiotics for the treatment of broncho-pulmonary processes exacerbations at children with chronic P. aeruginosa cystic fibrosis. Intravenous use of antibiotics out of hospital for the treatment of the children with cystic fibrosis is clinically effective, and is economically and psychologically reasonable. It should be used more widely in medical practice.

[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.

[Immunologic monitoring of patients with cystic fibrosis: value of different laboratory findings]. / Immunologicheskii monitoring bol'nykh mukovistsidozom, znachenie razlichnykh laboratornykh pokazatelei.

Cystic fibrosis (CF) is a common, serious, and frequently fatal autosomal recessive genetic disorder associated with the poor function of chloride channels. Chronic endobronchial inflammation and bacterial infection are main causes of morbidity and mortality due to CF. The study dealt with a relationship between progression and inflammation markers. Twenty one CF children with acute pulmonary exacerbation were examined. The signs of peripheral blood inflammation (responses of lymphocytes to PHA and their sensitivity to the antiproliferative effect of glucocorticoids) and in situ inflammation markers (sputum elastase activity, IL-8 and TNF-alpha, and protein concentrations in the same sputum specimens). These laboratory findings were used to calculate a "laboratory index" (LI). The clinical status of each patient was evaluated with a "clinical index" (CI), a parameter that includes respiratory secretion cultures, pulmonary function test results, nutritional status, and the presence of disease-related complications. There was a positive linear correlation between LI and CI. The presence of P. aeruginosa was strongly associated with the changes of inflammatory markers. CF patients with prolonged P. aeruginosa infection demonstrated extremely enhanced elastase activity and elevated amounts of sputum IL-8 and TNF-alpha as compared to uninfected subjects. The lung elastase activities, sputum protein contents, and TNF-alpha levels in individuals with short-term colonization were at or below those without P. aeruginosa infection. In patients with or without short-term colonization, the normalization of laboratory parameters was strongly related to evident clinical improvement. At the same time, antibiotic treatment failed to suppress an excessive inflammatory response in the lungs of patients with prolonged P. aeruginosa infection. The importance of individual inflammation markers is discussed in the paper.

Inflammatory markers in cystic fibrosis patients with lung Pseudomonas aeruginosa infection.

Chronic endobronchial inflammation and bacterial infection are the main causes of morbidity and mortality in cystic fibrosis (CF), an autosomal recessive genetic disorder associated with improper function of chloride channels. Inflammation in CF lung is greatly amplified after Pseudomonas aeruginosa infection. In this study the relationship between P. aeruginosa status and inflammatory markers has been investigated. Seventeen CF children in acute lung exacerbation were examined. CF patients without P. aeruginosa infection were characterized by elevated activity of sputum elastase, reduced response of peripheral blood lymphocytes to PHA and significant resistance to the antiproliferative action of glucocorticoids. These parameters were normalized after antibiotic treatment. The patients with prolonged P. aeruginosa infection demonstrated extremely high levels of elastase activity and elevated amounts of sputum IL-8 and TNF-alpha. Although antibiotic treatment resulted in clinical improvement, it failed to suppress excessive immune response in the lung. The data indicate that CF patients with prolonged P. aeruginosa need the modified treatment, which should include immunomodulating drugs and protease inhibitors as well as antibacterial therapy.

[Use ciprofloxacin in children with mucoviscidosis]. / Opyt primeneniia tsiprofloksatsina u detei, bol'nykh mukovistsidozom.

Ciprofloxacin was used in treatment of 13 children aged 6 to 18 years with mucoviscidosis and exacerbation of the bronchopulmonary process. The dose of the drug was 20 to 30 mg/kg a day when administered orally or 15 mg/kg a day when administered at first intravenously and then orally. The treatment course averaged 14 days. The indications to the drug use were: severe processes of mucoviscidosis and chronic colonization of the bronchial mucosa and lung tissues with Pseudomonas aeruginosa (mucoid or nonmucoid form) sensitive to ciprofloxacin and resistant to other antibiotics. The trials showed that ciprofloxacin was highly efficient: the state of the patients improved and the inflammation index of the total blood count normalized. However, eradication of P. aeruginosa from the respiratory tracts was not observed. The drug allergy in 1 patient and a transient increase in the level of transaminases in 5 patients as the adverse reactions were recorded.

[Characteristics of intrarenal ampicillin distribution in chronic pyelonephritis in children]. / Osobennosti vnutripochechnogo raspredeleniia ampitsillina pri khronicheskom pielonefrite u detei.

Pharmacokinetics and characteristic features of ampicillin kidney distribution were studied in 21 children with chronic pyelonephritis without signs of renal insufficiency who had undergone urological operations. It was found possible to provide the antibiotic concentrations efficient against ampicillin sensitive and partially middle sensitive microorganisms in the renal parenchyma, pelvis wall, ureterocele, megaureter and urinary bladder. Ampicillin concentrations in tissues of the urinary system were shown to correspond to a higher extent to the concentrations attained in blood than those in urine.

[Method of determining the antibiotic concentration in biological material]. / K metodike opredeleniia kontsentratsii nekotorykh antibiotikov v biologicheskom materiale.

To lower the role of natural antibacterial activity of biological substrates from humans and laboratory animals in microbiological assay of bleomycin (bleomycin) with B. subtilis ATCC 6633 as the test-microbe, it was suggested to increase the procedure sensitivity by using the medium modification. In determining concentrations of aminoglycosides by the agar diffusion method the use of B. pumilus NCTC 8241 as the test-microbe is recommended.

[Microflora study and the pharmacokinetic characteristics of gentamycin in children with mucoviscidosis]. / Izuchenie mikroflory i osobennostei farmakokinetiki gentamitsina u detei, bol'nykh mukovistsidozom.

P. aeruginosa and Staphylococcus were isolated respectively from 46.7 and 30.9 per cent of the children with mucoviscidosis. Proteus, Klebsiella and hemophilic bacilli were isolated from 11.2, 2.8 and 2.8 per cent of the patients respectively. All the isolates of Proteus, 95.8 per sent of the staphylococcal strains and 16.7 per cent of the P. aeruginosa strains were highly sensitive to gentamicin. No gentamicin resistant strains were detected. The studies on the gentamicin pharmacokinetics showed that the maximum levels of the drug in the blood and sputum of the children treated with the antibiotic injected intramuscularly in doses of 1.2 and 2-2.5 mg/kg were attained in 1 hour. The concentration of gentamicin in the sputum amounted to 60-80 per cent of its concentration in the blood serum. Within 11 hours the antibiotic sputum levels were higher than the MIC for the organisms sensitive to gentamicin. After a single endobronchial administration of the drug it was detected in the sputum for 6 days in concentrations exceeding the MIC for the moderately sensitive strains.