Таргетная терапия муковисцидоза при генотипе F508del/F508del

Targeted therapy for cystic fibrosis (CF) is a novel approach to CF treatment that can restore and potentiate CFTR channel activity. Lumacaftor/ivacaftor combination therapy is related to significant clinical and functional benefits in CF patients who are homozygous for F508del CFTR mutation. The authors described a case of effective treatment with lumacaftor/ivacaftor combination in a homozygous F508del CF patient with severe lung disease. This experience demonstrates an urgent need to make this pathogenic treatment available for patients with this genotype in Russia.Таргетная терапия муковисцидоза (МВ), при которой восстанавливается функция белка муковисцидозного трансмембранного регулятора (Cystic Fibrosis Transmembrane conductance Regulator – CFTR), является важнейшим достижением в лечении больных МВ. Убедительные успехи отмечены при применении комбинации «потенциатора» хлорного канала CFTR ивакафтора (Калидеко) и «корректора» люмакафтора. Представлен клинический случай успешного лечения препаратом ивакафтор / люмакафтор у пациентки с генотипом F508del/F508del (p.Phe508del, c.1521_1523delCTT, or c.1521_1523del or 1653delCTT) и тяжелым течением МВ. Этот опыт свидетельствует о необходимости активного распространения в России патогенетического лечения пациентов с аналогичным генотипом

Incidence of SARS-CoV-2 in people with cystic fibrosis in Europe between February and June 2020

Background Viral infections can cause significant morbidity in cystic fibrosis (CF). The current Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic could therefore have a serious impact on the health of people with CF (pwCF). Methods We used the 38-country European Cystic Fibrosis Society Patient Registry (ECFSPR) to collect case data about pwCF and SARS-CoV-2 infection. Results Up to 30 June 2020, 16 countries reported 130 SARS-CoV-2 cases in people with CF, yielding an incidence of 2.70/1000 pwCF. Incidence was higher in lung-transplanted patients (n=23) versus non-transplanted patients (n=107) (8.43 versus 2.36 cases/1000). Incidence was higher in pwCF versus the age-matched general population in the age groups <15, 15-24, and 25-49 years (p<0.001), with similar trends for pwCF with and without lung transplant. Compared to the general population, pwCF (regardless of transplantation status) had significantly higher rates of admission to hospital for all age groups with available data, and higher rates of intensive care, although not statistically significant. Most pwCF recovered (96.2%), however 5 died, of whom 3 were lung transplant recipients. The case fatality rate for pwCF (3.85%, 95% CI: 1.26-8.75) was non-significantly lower than that of the general population (7.46%; p=0.133). Conclusions SARS-CoV-2 infection can result in severe illness and death for pwCF, even for younger patients and especially for lung transplant recipients. PwCF should continue to shield from infection and should be prioritized for vaccination

Dornase alfa in the treatment of COVID-19: Destruction of neutrophil extracellular traps

In the severe or fatal course of COVID-19, rapid virus replication gives rise to exuberant inflammatory response including cytokine storm, characterized by elevated plasma concentrations of proinflammatory cytokines that support neutrophil activity and stimulate endothelial cells. Рost mortem examinations confirm these data, indicating extensive neutrophil infiltration and accumulation of neutrophil extracellular traps (NET)s in lung tissues of patients who have died from COVID-19. In this paper the possibility of therapy with dornase-alfa in COVID-19 patients is discussed. Designed to treat cystic fibrosis lung disease, this drug can reduce neutrophil activity, slow down the NET release and accelerate the NET clearance in the airways of COVID-19 patients. The authors also present the protocol of COVID-19 therapy with dornase-alfa produced by Russian manufacturer

Russian for Business Communication Textbooks in the University Information and Educational Environment

The article is devoted to the analysis of the didactic potential of multi-format textbooks on Russian for business communication (RBC) for linguo-communicative training of specialists in the areas of economics and management, receiving professional education in the universities of the Russian Federation. The study of the research source base, including theoretical and practical works, regulations and reference literature, allowed the authors to focus the methodological search on the design of an integrally whole and systemic flexible linguodidactic model of teaching RBC to Russian and foreign students regarding the full-time, distance or mixed formats of the pedagogical process and relying on printed and electronic educational tools as essential components of the university information and educational environment (real and virtual). The authors considered the subject contents of the RBC course and the ways of its presentation in various textbooks in the dialectical relationship of invariant and variable components as a condition for building a technological and adaptive educational model that ensures the interconnected formation of the entire complex of professional-communicative, sociocultural and intellectual-creative qualities of the specialist’s language personality. The permanent contents and didactic variability of RBC teaching aids for various contingents and various educational conditions are presented in the article using the example of author’s teaching aids created by the teachers of Bauman Moscow State Technical University (Russia, Moscow) and South-West State University (Russia, Kursk). The validity of the model realized in these manuals is confirmed by the practice of its use in the linguoeducational process at these universities

Timing of Ice Retreat Determines Summer State of Zooplankton Community in the Ob Estuary (the Kara Sea, Siberian Arctic)

In the estuaries of large Siberian rivers, ice coverage and the timing of ice retreat have varied in recent decades under the ongoing climate change. The seasonal development and functioning of the mesozooplankton community depend to a great extent on the timing of ice retreat. In the Arctic estuaries, the response of zooplankton to the timing of ice melt remains unclear. An earlier ice retreat was suggested to result in an advanced seasonal development of zooplankton, and higher biomass and feeding rates. Zooplankton composition, biomass, demography and grazing (assessed with the gut fluorescent approach) were studied in the Ob Estuary in July 2019 (“typical” ice retreat time). The obtained results were compared with the published data for July 2016 (ice retreat three weeks earlier). Zooplankton biomass in 2019 was considerably lower than in 2016, while species composition was similar; dominant populations were at an earlier stage of development. Herbivorous feeding of the dominant copepod, Limnocalanus macrurus, was also lower in 2019. The consequences of an earlier ice melt and increased temperature on seasonal population dynamics of the dominant brackish-water species are discussed. Our findings demonstrate that zooplankton communities in the Arctic estuaries are highly sensitive to the environmental changes associated with early sea-ice reduction

Airway and Lung Organoids from Human-Induced Pluripotent Stem Cells Can Be Used to Assess CFTR Conductance

Airway and lung organoids derived from human-induced pluripotent stem cells (hiPSCs) are current models for personalized drug screening, cell–cell interaction studies, and lung disease research. We analyzed the existing differentiation protocols and identified the optimal conditions for obtaining organoids. In this article, we describe a step-by-step protocol for differentiating hiPSCs into airway and lung organoids. We obtained airway and lung organoids from a healthy donor and from five donors with cystic fibrosis. Analysis of the cellular composition of airway and lung organoids showed that airway organoids contain proximal lung epithelial cells, while lung organoids contain both proximal and distal lung epithelial cells. Forskolin-induced swelling of organoids derived from a healthy donor showed that lung organoids, as well as airway organoids, contain functional epithelial cells and swell after 24 h exposure to forskolin, which makes it a suitable model for analyzing the cystic fibrosis transmembrane conductance regulator (CFTR) channel conductance in vitro. Thus, our results demonstrate the feasibility of generating and characterizing airway and lung organoids from hiPSCs, which can be used for a variety of future applications

Health Characteristics of Patients with Cystic Fibrosis whose Genotype Includes a Variant of the Nucleotide Sequence c.3140-16T&gt;A and Functional Analysis of this Variant

Cystic fibrosis (CF) is the most common monogenic autosomal recessive disease, associated with pathogenic variants in the CFTR gene. The splicing variant c.3140-16T&gt;A (3272-16T&gt;A) has been described previously and, according to the Russian CF Patients Registry, occurs with a frequency of 0.34%. The phenotypic features of CF patients with the c.3140-16T&gt;A variant were compared with those of patients with the genotype F508del/F508del. Patients with the allele c.3140-16T&gt;A had higher average age and age at diagnosis, and the allele was present in a greater proportion of adults. Patients carrying the c.3140-16T&gt;A allele were characterised by better physical development indicators, both in adults and in children, had preserved pancreatic function, as well as the absence of a number of complications, and required pancreatic enzyme replacement therapy less often than patients with the F508del/F508del genotype. Sweat test values also were lower in patients with the c.3140-16T&gt;A genotype. According to the results of clinical and laboratory studies, the phenotype of patients with the genetic variant c.3140-16T&gt;A can be considered “mild”. Functional CFTR protein activity in the presence of c.3140-16T&gt;A was evaluated using intestinal current measurements (ICM) and the forskolin-induced swelling assay on organoids obtained from patients’ rectal biopsies. c.3140-16T&gt;A had high residual CFTR channel activity and was amenable to effective pharmacological correction with thea VX-770 potentiator. To evaluate the effect of the variant on CFTR pre-mRNA splicing we performed a minigene assay, as well as RT-PCR analysis of RNA isolated from the nasal epithelium and rectal biopsy of patients. We showed that the c.3140-16T&gt;A variant creates a novel acceptor AG dinucleotide within CFTR intron 19, resulting in a 14-nucleotide extension of exon 20. This frameshift produces a premature termination codon and triggers mRNA degradation by the nonsense-mediated decay (NMD) mechanism. Moreover, we observed that the c.3140-16T&gt;A allele could produce a residual amount of normally spliced transcript, thus explaining the patient’s mild phenotype

Targeted sequencing reveals complex, phenotype-correlated genotypes in cystic fibrosis

Abstract Background Cystic fibrosis (CF) is one of the most common life-threatening genetic disorders. Around 2000 variants in the CFTR gene have been identified, with some proportion known to be pathogenic and 300 disease-causing mutations have been characterized in detail by CFTR2 database, which complicates its analysis with conventional methods. Methods We conducted next-generation sequencing (NGS) in a cohort of 89 adult patients negative for p.Phe508del homozygosity. Complete clinical and demographic information were available for 84 patients. Results By combining MLPA with NGS, we identified disease-causing alleles in all the CF patients. Importantly, in 10% of cases, standard bioinformatics pipelines were inefficient in identifying causative mutations. Class IV-V mutations were observed in 38 (45%) cases, predominantly ones with pancreatic sufficient CF disease; rest of the patients had Class I-III mutations. Diabetes was seen only in patients homozygous for class I-III mutations. We found that 12% of the patients were heterozygous for more than two pathogenic CFTR mutations. Two patients were observed with p.[Arg1070Gln, Ser466*] complex allele which was associated with milder pulmonary obstructions (FVC 107 and 109% versus 67%, CI 95%: 63-72%; FEV 90 and 111% versus 47%, CI 95%: 37-48%). For the first time p.[Phe508del, Leu467Phe] complex allele was reported, observed in four patients (5%). Conclusion NGS can be a more information-gaining technology compared to standard methods. Combined with its equivalent diagnostic performance, it can therefore be implemented in the clinical practice, although careful validation is still required

On Burkholderiales order microorganisms and cystic fibrosis in Russia

Abstract Background Microbes infecting cystic fibrosis patients’ respiratory tract are important in determining patients’ functional status. Representatives of Burkholderiales order are the most dangerous. The goal of our investigation was to reveal the diversity of Burkholderiales, define of their proportion in the microbiome of various parts of respiratory tract and determine the pathogenicity of the main representatives. Results In more than 500 cystic fibrosis patients, representing all Federal Regions of Russia, 34.0% were infected by Burkholderia cepacia complex (Bcc), 21.0% by Achromobacter spp. and 12.0% by Lautropia mirabilis. B. cenocepacia was the most numerous species among the Bcc (93.0%), and A. ruhlandii was the most numerous among Achromobacter spp. (58.0%). The most abundant genotype in Bcc was sequence type (ST) 709, and in Achromobacter spp. it was ST36. These STs constitute Russian epidemic strains. Whole genome sequencing of strains A. ruhlandii SCCH3:Ach33–1365 ST36 and B. cenocepacia GIMC4560:Bcn122 ST709 revealed huge resistomes and many virulence factors, which may explain the difficulties in eradicating these strains. An experience of less dangerous B. cenocepcia ST710 elimination was described. Massively parallel sequencing of 16S rDNA amplicons, including V1-V4 hypervariable regions, was used to definite “healthy” microbiome characteristics. Analysis of maxillary sinus lavage of 7 patients revealed infection with Proteobacteria of the same ST as pathogens from sputum, suggesting that the maxillary sinus is a source of infection in cystic fibrosis patients. Conclusions Characterization of the Russian epidemic bacterial strains in the sputum and sinuses of cystic fibrosis patients have better defined the importance of Burkholderiales bacteria. This information may aid in the development of effective approaches for treatment of this disease