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.

[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).

[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.

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.

Persistence of human papillomavirus type 6e in adult multiple laryngeal papilloma and the counterpart false cord of an interferon-treated patient.

Human papillomavirus (HPV) type 6e DNA was detected in a laryngeal papilloma, in a papilloma recurring after an interval of six years and in the counterpart clinically-normal false cord of the same patient. No detectable subgroup change of HPV DNA occurred during the clinical course, which included therapy by CO2 laser followed by intramuscular alpha-interferon injection. The data demonstrated a single subtype of HPV to have persisted, and the prevention of a recurrence of laryngeal papilloma during our patient's long clinical course to have failed, in spite of the interferon treatment.

Epstein-Barr virus DNA detection in gingival tissues of patients undergoing surgical extractions.

The main oral manifestation of Epstein-Barr virus (EBV) infection is hairy leukoplakia, a lesion associated with the acquired immunodeficiency syndrome (AIDS) and occasionally in other immunocompromised patients. However, the recent literature describes the presence of viral genome in clinically normal oral tissues. The purpose of this work was to investigate these occult EBV infections in gingival epithelium. The Southern blot method with 32P-radiolabelled DNA probes under stringent conditions was applied to 20 interproximal gingival papillae specimens and revealed homologous EBV sequences in 4 of 10 AIDS patients as well as in 4 of 10 HIV negative patients. In order to determine whether EBV has a predilection for the gingival tissues, samples of nasal, laryngeal and oral mucosa, other than gingival mucosa, were collected from 10 HIV-negative patients undergoing surgical treatment for a variety of clinical conditions. None of these extra-periodontal mucosal specimens contained homologous EBV DNAs, except an edentulous palatal gingival specimen. With the present detection of EBV DNAs in the gingival tissues of patients undergoing surgical extractions, it would be of interest to investigate more systematically these subclinical infections in order to determine their exact implications in oral disease.

Southern blot detection of human papillomaviruses (HPVs) DNA sequences in gingival tissues.

The highly sensitive and specific methods of molecular biology emphasize the frequency of subclinical infections in the genital tract tissues by the human papillomaviruses (HPVs). The purpose of this work was to investigate occult viral infections by the HPV type 6, 11, 16, and 18 in the gingival tissues. The Southern blot method with 32P-radiolabeled DNA probes applied under stringent conditions to 20 interproximal gingival papilla specimens revealed homologous viral sequences in 1 of 6 cases of adult periodontitis (HPV 16), 1 of 2 cases of rapidly progressive periodontitis (RPP) (HPV 6/HPV 11), 2 of 2 cases of acute gingivitis in psychiatric institutionalized patients (HPV 6; HPV 6/HPV 11), and 2 of 10 cases of acute gingivitis in AIDS patients (HPV 6/HPV 11/HPV 16; HPV 6). No periodontal or extra-periodontal specimen hybridized with the HPV 18 probe. Simultaneous hybridization with two or three HPV types was common (3/6 cases). The present detection of HPV 6, 11, 16 DNAs or related-DNAs in periodontal tissues without obvious clinical signs of viral infection suggests that the gingival epithelium may act as a reservoir.

Laryngeal candidiasis. Report of seven cases and review of the literature.

Although infections due to Candida have become increasingly recognized in recent years, laryngeal candidiasis remains a poorly described and infrequently diagnosed manifestation of mucous membrane candidal infection. Seven cases of isolated laryngeal candidiasis (ILC) have been identified at our institution during the past eight years (one before and six after death). Clinical, laboratory, and histopathologic findings from those seven cases, as well as from 12 additional cases reported in the literature, are reviewed. When hoarseness and dysphagia occur in patients with significant underlying disease who are receiving broad-spectrum antimicrobic therapy, a diagnosis of ILC should be considered. The diagnostic procedure of choice is indirect laryngoscopy with specimens submitted for culture and histopathologic study. On confirmation of the diagnosis, amphotericin B is the recommended therapy. Early treatment may limit morbidity and prevent systemic candidal dissemination.