Chronic obstructive pulmonary disease upper airway microbiome is associated with select clinical characteristics.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an inflammatory lung disorder associated with lung microbiome dysbiosis. Although the upper airway microbiome is the source of the lung microbiome, the relationships between the oral, nasal, and sputum microbiota are incompletely understood. Our objective was to determine features that differentiate the oral, nasal, and sputum microbiome among subjects with stable COPD. METHODS: We recruited 15 current or former smokers to provide oral and sputum samples on day 1. On day 2, another oral sample and a nasal sample were obtained. Each sample and control underwent DNA extraction, 16S V4 rRNA amplification, 16S V4 sequencing, and qPCR of 16S rRNA. Data were analyzed using dada2 and R. RESULTS: Most (14 of 15) subjects were male with a mean age of 65.2. One subject had no pulmonary obstruction, while 5 had mild COPD, 7 had moderate COPD, and 2 had severe COPD. Three subjects (20%) were current tobacco users and 2 subjects (13%) used inhaled corticosteroids (ICS). Subjects had a mean of 49.1 pack-years of tobacco exposure. Bacterial biomass was associated with anatomic site, but no differences in biomass were observed with age, FEV1 percent predicted (FEV1pp), ICS use, smoking status, or edentulous state. Shannon index was associated with site (lower nasal diversity than oral and sputum diversity, p<0.001), but not age, ICS use, FEV1pp, tobacco use, or edentulous state. ß-diversity was illustrated by principal coordinate analysis using Bray-Curtis dissimilarity and PERMANOVA analyses, showing sample clustering by anatomic site (p = 0.001) with nasal samples forming a cluster separate from the combined oral wash samples and sputum samples. Clustering was also observed with ICS use (p = 0.029) and edentulous state (p = 0.019), while FEV1pp and current tobacco use were not significant. In an amplicon sequencing variant (ASV)-level analysis of oral samples using a linear regression model with Benjamini-Hochberg correction at an FDR<0.10, 10 ASVs were associated with age while no ASVs were associated with FEV1pp or smoking status. Sputum sample analysis demonstrated that 51 ASVs (25 unique genera) were associated with age, 61 ASVs (32 genera) were associated with FEV1pp, and no ASVs were associated with smoking status. In a combined dataset, the frequent exacerbator phenotype, rather than ICS use, was associated with decreased sputum Shannon diversity. CONCLUSIONS: Among the upper airway microbiota of COPD subjects, anatomic site was associated with bacterial biomass, Shannon diversity, and ß-diversity. ICS use and edentulous state were both associated with ß-diversity. Age was associated with taxa relative abundance in oral and sputum samples, while FEV1pp was associated with taxa relative abundance in sputum samples only.

Influence of subglottic secretion drainage on the microorganisms of ventilator associated pneumonia: A meta-analysis for subglottic secretion drainage.

BACKGROUND: The influence of the subglottic secretion drainage (SSD) on the microorganisms of ventilator associated pneumonia (VAP) is still unclear.A meta-analysis focusing on the influence of the SSD on the microorganisms of VAP. METHODS: A comprehensive search was conducted through the online studies of PubMed, Embase, Cochrane Library, Google scholar, CNKI (China National Knowledge Infrastructure), and VIPI (Database for Chinese Technical Periodicals) using specific search terms.Included studies were randomized controlled trials (RCTs) that compare the microorganisms of VAP between SSD and standard endotracheal tube care in mechanically ventilated adults. RESULTS: Nine RCTs were eligible. There was no significant difference in the rate of VAP caused by nonfermentative bacteria and enterobacteria between SSD group and control group (OR = 0.73, 95%CI, 0.53-1.01; P = .06). The episodes of VAP caused by Gram-positive cocci and Haemophilus influenzae organisms were lower in the SSD group (OR = 0.29, 95%CI, 0.18-0.48; P<0.00001). Less mean volume of SSD daily was observed in VAP group (OR = -16.97, 95%CI, -29.87-4.08; P = .010). CONCLUSION: We found SSD to be associated with significant decreases in VAP caused by Gram-positive cocci and H influenzae organisms but no significant differences in VAP caused by nonfermentative bacteria and enterobacteria. Less mean volume of SSD daily was observed in VAP group.

[The treatment of laryngomycosis]. / Lechenie laringomikoza.

The objective of the present study was to elaborate the therapeutic algorithm for the treatment of laryngomycosis. We have examined a total of 430 patients suffering from chronic laryngitis including 100 ones (23.2%) having the fungal flora. Mycosis was diagnosed by the microscopic study of the stained preparations with the application of various techniques; moreover, cultivation in elective nutrient media was used. The yeast-like fungi of the genus Candida were identified in 98 (98%) and mold fungi of the genus Aspergillus in the remaining 2 (2%) patients. All these 100 patients were given the antifungal treatment. 98 of them presenting with candidal laryngitis were allocated to three groups. Group A was comprised of 33 patients who received the local treatment alone, group B contained 31 patients given only systemic therapy, and group C included 34 patients undergoing the combined treatment with the use of the medications possessed of both the local and systemic actions. The best clinical results were obtained in the patients of the latter group in which the therapeutic efficiency proved to be as high as 79.4%. It is concluded that all the patients suffering from laryngomycosis are in need of the combined treatment designed to eradicate the causative agent, restore the vocal function, and achieve the stable remission of the chronic inflammatory process in the larynx. The combined treatment with the antimycotic medications exhibiting both the general systemic and local activities during 3 weeks appears to be the optimal therapeutic modality for the management of laryngomycosis. Of special importance is the dynamic follow-up of the treated patients including the control examinations in the course of the treatment (days 7, 14, and 21) to be followed by the mycological study after the termination of therapy and its repetition every 3 months during the subsequent period.

[Microflora of laryngeal mucosa in the healthy subjects and the patients presenting with non-specific post-intubation laryngeal granuloma].

This comparative study was designed to determine the qualitative and quantitative composition of microflora of laryngeal mucosa in the healthy subjects and the patients presenting with non-specific post-intubation laryngeal granuloma. The secondary objective was to measure the sensitivity of the isolated microorganisms to antibiotic therapy. The quantitative characteristic and the species composition of the microbial associations in laryngeal mucosa were investigated in 40 healthy subjects and 72 patients presenting with non-specific post-intubation laryngeal granuloma. The mean age of the participants of the study varied from 31 to 69 years. The total microbial population density in the healthy subjects was 2.672±614 CFU/ml compared with 21.514±2.382 CFU/ml in the patients. The healthy subjects were found to carry streptococci in 87.5% of the cases, staphylococci and neisseria in 60.0% and 42.5% of the cases respectively. As many as 77.8%, 83.3%, and 90.3% of the patients with non-specific post-intubation laryngeal granuloma were carriers of these microorganism respectively. The dominant microorganisms in the mucous membrane of the larynx in these patients were N. flavescence (17.6%), S. aureus (9.01%), S. pneumonia (7.3%), and S. pyogenes (6.0%).The staphylococci isolated from the mucous membrane of the larynx of the patients presenting with non-specific post-intubation laryngeal granuloma exhibited the highest sensitivity to cephalosporin antibiotics (cefuroxime, cefprome or ceftriaxone) and fluoroquinolone (levofloxacin) antibiotics (100, 100, 91.7 and 96.7% respectively).

Pneumococcal phosphoglycerate kinase interacts with plasminogen and its tissue activator.

Streptococcus pneumoniae is not only a commensal of the nasopharyngeal epithelium, but may also cause life-threatening diseases. Immune-electron microscopy studies revealed that the bacterial glycolytic enzyme, phosphoglycerate kinase (PGK), is localised on the pneumococcal surface of both capsulated and non-capsulated strains and colocalises with plasminogen. Since pneumococci may concentrate host plasminogen (PLG) together with its activators on the bacterial cell surface to facilitate the formation of plasmin, the involvement of PGK in this process was studied. Specific binding of human or murine PLG to strain-independent PGK was documented, and surface plasmon resonance analyses indicated a high affinity interaction with the kringle domains 1-4 of PLG. Crystal structure determination of pneumococcal PGK together with peptide array analysis revealed localisation of PLG-binding site in the N-terminal region and provided structural motifs for the interaction with PLG. Based on structural analysis data, a potential interaction of PGK with tissue plasminogen activator (tPA) was proposed and experimentally confirmed by binding studies, plasmin activity assays and thrombus degradation analyses.

Polysaccharide-protein conjugate vaccination induces antibody production but not sustained B-cell memory in the human nasopharyngeal mucosa.

Colonization of the nasopharyngeal mucosa by meningococcus and other polysaccharide (PS)-encapsulated bacteria precedes invasion. PS-conjugate vaccines induce PS-specific B-cell memory (B(MEM)) and also prevent colonization, thus blocking person-to-person transmission, generating herd protection. However, in isolation the B(MEM) are unable to sustain immunity. Furthermore, the duration of herd protection the vaccines induce appears limited. We demonstrate that, despite the persistence of PS-specific B(MEM), the population is not maintained within the nasopharynx. Although booster immunization results in the transient appearance of PS-specific B(MEM) within the mucosa, this reflects the re-circulation of systemic B(MEM) through the site rather than the generation of resident mucosal B(MEM). The induction of sustained PS-specific B(MEM) in the nasopharynx would allow the population to be activated by colonization, thus inhibiting subsequent invasion. It would also be expected to boost local mucosal immunity, thus extending herd protection. Strategies to generate PS-specific B(MEM) in the mucosa warrant further investigation.

Absence of Helicobacter pylori in healthy laryngeal mucosa.

OBJECTIVES: To evaluate the presence of Helicobacter pylori in healthy laryngeal mucosa. DESIGN: Prospective analysis of Helicobacter pylori colonisation in healthy laryngeal mucosa, using the 13C urea breath test and polymerase chain reaction analysis. SUBJECTS: Twenty randomly chosen men (28-78 years) without laryngeal pathology or gastrointestinal disease were investigated. All subjects were scheduled for elective operative procedures, under general, endotracheal anaesthesia. Cytobrush samples were taken for Helicobacter pylori DNA detection. Nested polymerase chain reaction testing was performed on DNA solutions using two primer pairs from the urease A gene of the Helicobacter pylori genome. The 13C urea breath test was performed on two exhalation samples. RESULTS: Eight (40 per cent) of the subjects were positive for urease on urea breath testing; none were positive for Helicobacter pylori DNA on polymerase chain reaction testing. CONCLUSION: Based on these results, we do not consider Helicobacter pylori to be a normal constituent of healthy laryngeal microflora.

Biofilm accumulation on endotracheal tubes following prolonged intubation.

OBJECTIVE: To demonstrate that patients who have been intubated for prolonged periods of time will have an increased likelihood of developing bacterial biofilm on their endotracheal tubes. METHODS: We collected endotracheal tubes from patients at the time of extubation, and analysed representative sections with scanning electron microscopy for morphologic evidence of biofilms. RESULTS: From September 2007 to September 2008, 32 endotracheal tubes were analysed with electron microscopy. Patients who had been intubated for 6 days or longer had a significantly higher percentage of endotracheal tubes that exhibited bacterial biofilms, compared with patients intubated for less than 6 days (88.9 versus 57.1 per cent, p = 0.0439). CONCLUSIONS: Longer duration of intubation is associated with a higher incidence of bacterial biofilm. Further research is needed to link the presence of bacterial biofilms to acquired laryngotracheal damage.

[The specific features of microbiocenoses in children living under conditions of anthropogenic pressing].

The specific features of enteric and nasopharyngeal microbiocenoses and the species composition of bifidobacteria have been studied in children living in the industrial towns of the Irkutsk Region under the existing anthropogenic load. Ambient air pollution is characterized and a presumptive human health risk assessed.

[Characteristics of microflora of laryngeal mucosa in healthy subjects and patients with cicatrical stenosis of the larynx].

The authors present data on the occurrence, density, and species composition of microorganisms isolated from pharyngeal mucosa of 40 healthy subjects and 55 patients with cicatrical stenosis of the larynx. The total density of microbial populations in control subjects was estimated at 2,672+/-614 CFU/ml compared with 17,510+/-1,983 CFU/ml in patients with cicatrical stenosis of the larynx unrelated to malignant neoplastic growth and 19,375+/-2,103 CFU/ml in patients who had experienced surgical intervention for the treatment of malignancy. A total of 312 microbial strains were isolated and identified. Coccal bacteria were the predominant group (streptococci-74 strains, neisseria-72, staphylococci-62, micrococci-12). Collectively, they comprised 70.5% of all isolated strains representing autochthonous microflora. Corynebacteria, moraxellas, and enterobacteria (transitory forms) were rare. The occurrence of neisseria, staphylococci and fungi (genus Candida) in patients with cicatrical stenosis of the larynx was significantly higher than in the healthy subjects. Moreover, the patients developed dysbacteriosis with the predominance of N. flavescens, S. aureus, S. epidermidis, S. alpha-haemoliticus, and Candida sp. It is concluded that elucidation of the microbial landscape in patients with cicatrical stenosis of the larynx is of importance for planning prevention and management of pyoseptic complications in the course of surgical treatment.

An interesting entity mimicking malignancy: laryngeal candidiasis.

Primary candidiasis in the laryngeal mucosa is rare but has many predisposing factors, in particular immune insufficiency and long-term antibiotic use. This study involved a reassessment of 530 laryngeal biopsy specimens originally examined between 2004 and 2009, from which 11 cases of hyperplastic laryngeal candidiasis were identified according to their clinical and histopathological features. Among possible predisposing factors, immune insufficiency was identified in three patients, there was long-term antibiotic use in two cases, use of inhaled corticosteroids in four cases, smoking in eight cases and reflux symptoms in four cases. Dysplasia as well as hyperplasia was seen in two cases. Endolaryngeal lesion excision was performed in eight cases and biopsy leading to radical surgery was done in three. In clinical follow-up no lesions were observed in nine cases while two had new leukoplakic lesions.

Physiologic cold shock increases adherence of Moraxella catarrhalis to and secretion of interleukin 8 in human upper respiratory tract epithelial cells.

Moraxella catarrhalis, a major nasopharyngeal pathogen of the human respiratory tract, is exposed to rapid and prolonged downshifts of environmental temperature when humans breathe cold air. In the present study, we show that a 26 degrees C cold shock up-regulates the expression of UspA1, a major adhesin and putative virulence factor of M. catarrhalis, by prolonging messenger RNA half-life. Cold shock promotes M. catarrhalis adherence to upper respiratory tract cells via enhanced binding to fibronectin, an extracellular matrix component that mediates bacterial attachment. Exposure of M. catarrhalis to 26 degrees C increases the outer membrane protein-mediated release of the proinflammatory cytokine interleukin 8 in pharyngeal epithelial cells. Furthermore, cold shock at 26 degrees C enhances the binding of salivary immunoglobulin A on the surface of M. catarrhalis. These data indicate that cold shock at a physiologically relevant temperature of 26 degrees C affects the nasopharyngeal host-pathogen interaction and may contribute to M. catarrhalis virulence.

Laryngopharyngeal reflux finding scores correlate with gastroesophageal reflux disease and Helicobacter pylori expression.

CONCLUSION: The expression of Helicobacter pylori (HP) positivity and degree of gastroesophageal reflux disease (GERD) correlate with laryngopharyngeal reflux (LPR). HP positivity and degree of GERD were more adverse in patients with a reflux finding score (RFS) of 7 or more. OBJECTIVE: We aimed to investigate the relationship between RFS and inflammation of the lower part of the esophagus as well as RFS and HP infection. PATIENTS AND METHODS: Forty-five consecutive patients were analyzed prospectively. The degree of LPR was evaluated using the RFS method. The degree of GERD, lower esophageal mucosal inflammation, and antral HP positivity were evaluated using endoscopic surveys. RESULTS: The mean RFS of the whole population was 11.5+/-4.4. The mean RFS of patients who had lower esophageal mucosal inflammation was 7+/-0.1, 8.1+/-1.3, 13.9+/-3.7, and 16.6+/-3.5, for grades A, B, C and D, respectively. The RFS of patients according to HP expression was as follows: 7.2+/-0.4, 9.3+/-3.07, 12.7+/-3.16, and 17.8+/-2.1, for normal (score 0), score I, score II, and score III, respectively.

Investigation of Helicobacter pylori colonization in laryngeal neoplasia.

Helicobacter pylori has been investigated in several other organ systems and localizations such as the oral cavity, but has not been investigated extensively in squamous cell carcinoma of the larynx, a region that could be directly exposed to the bacterium by the oral route or gastro-esophageal reflux. Only serological studies are available regarding the relation between H. pylori and laryngeal cancer, yielding conflicting results. To our knowledge, there is no study investigating the presence of H. pylori in laryngeal squamous cell carcinoma tissue. The purpose of this study was to investigate the presence of H. pylori in laryngeal squamous cell carcinoma tissue and to investigate the possible role of this organism in the etiopathogenesis of laryngeal cancer. Specimens from 50 patients with laryngeal cancer who underwent total or partial laryngectomy between March 1999 and December 2002 were examined by histopathological and immunohistochemical methods to detect H. pylori. The presence of H. pylori was also investigated histopathologically in 50 benign laryngeal biopsy specimens. In our study, we demonstrated that H. pylori was not present in laryngeal squamous cell carcinoma tissue or in the benign lesions. We could not find any evidence indicating that H. pylori played a role at the tissue level in the pathogenesis of laryngeal carcinoma.

[Infection control in neutropenia induced by high-dose cytarabine chemotherapy].

A high-dose cytarabine (Cylocide; Ara-C: HDAC) chemotherapy has been successfully used as a postremission consolidation therapy for acute myeloid leukemia (AML). Although this chemotherapy has been estimated to cause severe myelosuppression, there has been no report about infection risk relating to HDAC chemotherapy. The purpose of this retrospective study is to evaluate the infection risk in AML patients treated with HDAC (n = 18) compared to those treated with standard-dose Ara-C (SDAC, n = 18). The mean duration of severe neutropenia (neutrophils < 500/microl) in HDAC group and SDAC was 14.8 days and 10.4 days, respectively, indicating a significant prolongation in the HDAC group (p < 0.05). The frequency of febrile neutropenia in the HDAC group tended to increase compared to that in the SDAC group (p = 0.093). The average days of usage of quinolone antimicrobial prophylaxis and aminoglycoside antibiotic injection in febrile neutropenia in the HDAC group were significantly longer than those of the SDAC group (quinolone; p < 0.01, aminoglycoside; p < 0.05). The frequency of Streptococcus infection isolated from pharyngeal mucus in the HDAC group was significantly higher than that in the SDAC group (100% versus 75%; p < 0.05). These results suggest that HDAC chemotherapy increased the infection risk compared to SDAC, and especially patients who received HDAC need a further prevention plan against gram-positive bacteria.

[Laryngeal infection by Rhodococcus equi in patient with AIDS]. / Laringitis por Rhodococcus equi en paciente con SIDA.

The Rhodococcus equi is an aerobic gram positive pleomorphic bacillus, that was isolated for the first time like a producer of bronchopneumonia in young horses. Every time more often, it is being recognized as a pathogen in humans, mainly in the immunodepressed population. We described a case, until now exceptional, of laryngeal infection by Rhodococcus equi in a patient with positive serology for the virus of the human immunodeficiency (HIV), and we reviewed some clinical and epidemiological characteristics of the infections by this germ. The treatment is riphampicine and/or erythromycin, being the prognosis bad, because usually they are immunodepressed patients.

Molecular determinants of the pathogenesis of disease due to non-typable Haemophilus influenzae.

Non-typable Haemophilus influenzae is a common commensal organism in the human upper respiratory tract and an important cause of localized respiratory tract disease. The pathogenesis of disease begins with bacterial colonization of the nasopharynx, a process that involves establishment on the mucosal surface and evasion of local immune mechanisms. Under the proper circumstances, the organism spreads contiguously to the middle ear, the sinuses, or the lungs, and then stimulates a brisk inflammatory response, producing symptomatic infection. In this review, we summarize our present understanding of the molecular determinants of this sequence of events. Continued investigation of the molecular mechanism of non-typable H. influenzae pathogenicity should facilitate development of novel approaches to the treatment and prevention of H. influenzae disease.

Clinical and endoscopic findings in the mucosae of the upper respiratory and digestive tracts in post-treatment follow-up of paracoccidioidomycosis patients.

Systematic examination of the upper respiratory and digestive tracts (URDT) was performed in a group of 80 paracoccidioidomycosis (PCM) patients submitted to post-treatment follow-up ranging from 8 months to 17 years. Mucosae of the URDT had been involved prior to specific treatment in 74 patients, distributed as follows: oropharynx, 50 (41 alone, 7 in association with the larynx, and 2 with the nasal mucosa); larynx, 30 (23 alone and 7 in association); and nasal mucosa, 3 (1 alone and 2 in association). Inactive lesions were observed in all the 50 patients with lesions of the oropharynx, 3 of whom with deforming scars (1 with retraction of the tongue and 2 with narrowing of the oral orifice). One case presented a destructive lesion, with perfuration of the palate. Of the other 46 cases, examination showed nacreous white striated scars which were nearly imperceptible in some cases and in others displayed partial retraction of anatomical structures without any alteration of their features. Patients presented a high rate of missing teeth. In 3 patients with involvement of the nasal mucosa, none of whom presented active PCM lesions, 2 still had nasal voices. In 30 patients with lesions of the larynx, 1 suffered a relapse of PCM and 2 developed epidermoid carcinoma. Of the other 27 cases, none of whom had active PCM lesions, 15 presented dysphonia, 3 were tracheotomized, and 9 were asymptomatic.