RESUMO
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.
Assuntos
Burkholderiales/genética , Burkholderiales/fisiologia , Fibrose Cística/microbiologia , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Feminino , Variação Genética , Humanos , Lactente , Masculino , Microbiota , Pessoa de Meia-Idade , Federação Russa , Adulto JovemRESUMO
BACKGROUND AND AIM: The order Burkholderiales became more abundant in the healthcare units since the late 1970s; it is especially dangerous for intensive care unit patients and patients with chronic lung diseases. The goal of this investigation was to reveal the real variability of the order Burkholderiales representatives and to estimate their phylogenetic relationships. METHODS: 16S rDNA and genes of the Burkholderia cenocepacia complex (Bcc) Multi Locus Sequence Typing (MLST) scheme were used for the bacteria detection. RESULTS: . A huge diversity of genome size and organization was revealed in the order Burkholderiales that may prove the adaptability of this taxon's representatives. The following variability of the Burkholderiales in Russian healthcare units has been revealed: Burkholderiaceae (Burkholderia, Pandoraea, and Lautropia), Alcaligenaceae (Achromobacter), and Comamonadaceae (Variovorax). The Burkholderia genus was the most diverse and was represented by 5 species and 16 sequence types (ST). ST709 and 728 were transmissible and often encountered in cystic fibrosis patients and in hospitals. A. xylosoxidans was estimated by 15 genotypes. The strains of first and second ones were the most numerous. CONCLUSIONS: Phylogenetic position of the genus Lautropia with smaller genome is ambiguous. The Bcc MLST scheme is applicable for all Burkholderiales representatives for resolving the epidemiological problems.
Assuntos
Infecções por Burkholderia/genética , Burkholderiaceae/genética , Filogenia , RNA Ribossômico 16S/genética , Infecções por Burkholderia/epidemiologia , Infecções por Burkholderia/microbiologia , Burkholderiaceae/patogenicidade , Variação Genética , Genótipo , Humanos , Especificidade da EspécieRESUMO
Using immunoblotting with recombinant recoverin as an antigen, we have examined 279 serum samples from individuals with small cell lung carcinoma (SCLC, 99 patients), non-small cell lung carcinoma (NSCLC, 44 patients), and non-malignant pulmonary disorders (86 patients) as well as sera from 50 healthy donors. Autoantibodies against recoverin (anti-Rc) were detected in sera from 15 patients with SCLC (15% of cases) and from 9 patients with NSCLC (about 20% of cases). Only two anti-Rc positive cases were detected in patients with non-malignant pulmonary disorders, while no such cases were found in healthy individuals. Immunohistochemical investigation of paraffin sections of 44 SCLC and 40 NSCLC tumors revealed recoverin-positive reaction in 30 SCLC (68%) and 34 NSCLC (85%) sections. Despite the high specificity (98%), the low sensitivity (less than 20%) does not allow serum anti-Rc to be considered as a valuable marker of lung cancer. However, taking into account the high occurrence of aberrant expression of recoverin in lung tumors, this PNA could be considered as a potential target for immunotherapy of lung cancer.
Assuntos
Antígenos de Neoplasias/análise , Autoanticorpos/análise , Biomarcadores Tumorais/análise , Proteínas de Ligação ao Cálcio/análise , Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma de Células Pequenas/imunologia , Proteínas do Olho , Lipoproteínas , Pneumopatias/imunologia , Neoplasias Pulmonares/imunologia , Proteínas do Tecido Nervoso , Idoso , Feminino , Hipocalcina , Humanos , Immunoblotting , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Recoverina , Sensibilidade e EspecificidadeRESUMO
The goal of the present study was to analyze serum and tumor tissue of a patient with non-small cell lung cancer (NSCLC) for the presence of autoantibodies against recoverin (anti-Rc) and recoverin expression, correspondingly. Using immunoblotting with recombinant recoverin as an antigen, we have detected anti-Rc in serum of the patient. At the same time, the patient did not manifest any signs of cancer-associated retinopathy (CAR). Polyclonal (monospecific) antibodies against recoverin used for immunohistochemical analysis of the patient's tumor revealed recoverin expression in the tumor sections. To our knowledge, this is the first case of the presence of serum anti-Rc in NSCLC patients in the absence of paraneoplastic retina degeneration.