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1.
Clin Lab ; 65(8)2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31414746

RESUMO

BACKGROUND: Foreign body aspiration is a rare entity in adults. We presented an adult case with recurrent pulmonary infection firstly misdiagnosed as tuberculosis, which proved as foreign body aspiration in the left main stem bronchus by bronchoscopy. METHODS: Appropriate laboratory tests are carried out. The chest CT scan and bronchoscopy were performed for diagnosis. RESULTS: Serum sedimentation was increased and tuberculosis antibody was positive. The chest CT scan showed left lung consolidation and small pleural exudate on the left side. Significant calcification can be seen near the left main bronchus. The bronchoscopy demonstrated plenty of yellow sputum in left main bronchus and a peanut shell completely obstructed the left main bronchus and peripheral granulation tissue hyperplasia. The peanut shell was removed and the left main trachea was unobstructed. CONCLUSIONS: When a patient has recurrent pulmonary infection, especially at the same site, physicians should pay attention to airway obstruction caused by foreign body, cancer and other causes of airway stenosis. Bronchoscopy is crucial for the ultimate diagnosis.


Assuntos
Sedimentação Sanguínea , Erros de Diagnóstico , Corpos Estranhos/diagnóstico , Pulmão/diagnóstico por imagem , Mycobacterium tuberculosis/imunologia , Tuberculose/diagnóstico , Idoso , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Brônquios/microbiologia , Brônquios/patologia , Broncoscopia , Humanos , Pulmão/microbiologia , Masculino , Mycobacterium tuberculosis/fisiologia , Tomografia Computadorizada por Raios X , Traqueia/microbiologia , Traqueia/patologia , Tuberculose/sangue , Tuberculose/microbiologia
2.
BMC Infect Dis ; 19(1): 598, 2019 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-31288744

RESUMO

BACKGROUND: In Japan and other countries, the number of patients with syphilis is increasing year by year. Recently, the cases of the pulmonary involvement in patients with secondary syphilis have been reported. However, it is still undetermined how to obtain a desirable specimen for a diagnosis of the pulmonary involvement, and how to treat it if not cured. CASE PRESENTATION: A 34-year-old man presented with cough and swelling of the right inguinal nodes. A physical examination revealed erythematous papular rash over the palms, soles and abdomen. A 4 cm mass in the right lower lobe of the lung was detected on computed tomography. He was diagnosed as having secondary syphilis, because he was tested positive for the rapid plasma reagin and Treponema pallidum hemagglutination assay. Amoxycillin and probenecid were orally administered for 2 weeks. Subsequently, rash and serological markers were improved, however, the lung mass remained unchanged in size. Transbronchial biopsy (TBB) confirmed the pulmonary involvement of syphilis using polymerase chain reaction techniques (tpp47- and polA-PCR). Furthermore, following surgical resection revealed the lung mass to be an abscess. CONCLUSIONS: To our knowledge, this is the first surgically treated case of a lung abscess caused by syphilis, which was diagnosed by PCR techniques in TBB. This report could propose a useful diagnostic method for the pulmonary involvement of syphilis.


Assuntos
Reação em Cadeia da Polimerase/métodos , Sífilis/diagnóstico , Adulto , Brônquios/microbiologia , Brônquios/patologia , Proteína C-Reativa/análise , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/metabolismo , Humanos , Masculino , Sífilis/microbiologia , Tomografia Computadorizada por Raios X , Treponema pallidum/genética , Treponema pallidum/isolamento & purificação
3.
Vet Microbiol ; 235: 80-85, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282382

RESUMO

Bovine respiratory disease complex is a major disease affecting the global cattle industry. Multiple infections by viruses and bacteria increase disease severity. Previously, we reported that bovine respiratory syncytial virus (BRSV) infection increases adherence of Pasteurella multocida to human respiratory and bovine kidney epithelial cells. To examine the interaction between the virus and bacteria in bovine respiratory cells, we generated respiratory epithelial cell lines from bovine trachea (bTEC), bronchus (bBEC), and lung (bLEC). Although all established cell lines were infected by BRSV and P. multocida susceptibility differed according to site of origin. The cells derived from the lower respiratory tract (bBEC and bLEC) were significantly more susceptible to BRSV than those derived from the upper respiratory tract (bTEC). Pre-infection of bBEC and bLEC with BRSV increased adherence of P. multocida; this was not the case for bTEC. These results indicate that BRSV may reproduce better in the lower respiratory tract and encourage adherence of bacteria. Thus, we identify one possible mechanism underlying severe pneumonia.


Assuntos
Coinfecção/veterinária , Células Epiteliais , Interações Microbianas , Infecções por Pasteurella/veterinária , Infecções por Vírus Respiratório Sincicial/veterinária , Animais , Complexo Respiratório Bovino/microbiologia , Complexo Respiratório Bovino/virologia , Brônquios/citologia , Brônquios/microbiologia , Brônquios/virologia , Bovinos , Linhagem Celular , Células Cultivadas , Coinfecção/microbiologia , Coinfecção/virologia , Citocinas/metabolismo , Células Epiteliais/microbiologia , Células Epiteliais/virologia , Pulmão/citologia , Pulmão/microbiologia , Pulmão/virologia , Infecções por Pasteurella/virologia , Pasteurella multocida/genética , Pasteurella multocida/isolamento & purificação , Infecções por Vírus Respiratório Sincicial/microbiologia , Vírus Sincicial Respiratório Bovino/genética , Vírus Sincicial Respiratório Bovino/isolamento & purificação , Traqueia/citologia , Traqueia/microbiologia , Traqueia/virologia
4.
Mycoses ; 62(10): 883-892, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31166635

RESUMO

The current knowledge of invasive Scopulariopsis/Microascus infection in lung transplantation has been derived from only four case reports. Although these fungi are uncommon compared with Aspergillus, they are highly resistant to the current antifungal agents, and the mortality is extremely high. To explore the risk factors, clinical manifestations, notable diagnostic characteristics and outcomes of positive Scopulariopsis/Microascus isolation in lung transplantation patients. We included all cases with positive Scopulariopsis/Microascus isolation from lower respiratory tracts or bronchial mucosa biopsies in our lung transplantation centre. Proven cases from the literature were added. Positive isolation occurred in 2% (3/157) in our centre. Four cases from the literature were added. The mortality could be considered as high as 80%, once the two cases of colonisation were excluded. The average interval between transplantation and positive isolation was 106 (19-131) days. A total of 57.1% of patients had experienced a combination of infection with Aspergillus or other fungi as well as long-term azole antifungal agent treatment before the positive isolation, which may be possible risk factors. The combination of micafungin, posaconazole and terbinafine may be an effective treatment. The peak time of positive isolation was consistent with that of some opportunistic pathogens, and the possible risk factors were the infection of other fungi as well as prior long-term azole antifungal administration. In addition to its high mortality, Scopulariopsis/Microascus was also highly resistant to common antifungal agents and the combination of two or three drugs for therapy was recommended.


Assuntos
Brônquios/microbiologia , Transplante de Pulmão/efeitos adversos , Pulmão/microbiologia , Micoses/diagnóstico , Micoses/patologia , Scopulariopsis/isolamento & purificação , Transplantados , Adulto , Idoso , Antifúngicos/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Micoses/epidemiologia , Micoses/microbiologia , Prevalência , Fatores de Risco , Análise de Sobrevida , Adulto Jovem
5.
Eur J Med Chem ; 175: 63-71, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31075609

RESUMO

In the frame of a research program aimed to explore the relationship between chirality of iminosugars and their therapeutic potential, herein we report the synthesis of N-akyl l-deoxyiminosugars and the evaluation of the anti-inflammatory properties of selected candidates for the treatment of Pseudomonas aeruginosa infections in Cystic Fibrosis (CF) lung disease. Target glycomimetics were prepared by the shortest and most convenient approach reported to date, relying on the use of the well-known PS-TPP/I2 reagent system to prepare reactive alkoxyalkyl iodides, acting as key intermediates. Iminosugars ent-1-3 demonstrated to efficiently reduce the inflammatory response induced by P. aeruginosa in CuFi cells, either alone or in synergistic combination with their d-enantiomers, by selectively inhibiting NLGase. Surprisingly, the evaluation in murine models of lung disease showed that the amount of ent-1 required to reduce the recruitment of neutrophils was 40-fold lower than that of the corresponding d-enantiomer. The remarkably low dosage of the l-iminosugar, combined with its inability to act as inhibitor for most glycosidases, is expected to limit the onset of undesired effects, which are typically associated with the administration of its d-counterpart. Biological results herein obtained place ent-1 and congeners among the earliest examples of l-iminosugars acting as anti-inflammatory agents for therapeutic applications in Cystic Fibrosis.


Assuntos
Antibacterianos/uso terapêutico , Fibrose Cística/complicações , Imino Açúcares/uso terapêutico , Infecções por Pseudomonas/complicações , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/isolamento & purificação , Animais , Antibacterianos/administração & dosagem , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Brônquios/imunologia , Brônquios/microbiologia , Brônquios/patologia , Relação Dose-Resposta a Droga , Humanos , Imino Açúcares/administração & dosagem , Imino Açúcares/química , Imino Açúcares/farmacologia , Inflamação/prevenção & controle , Concentração Inibidora 50 , Camundongos , Neutrófilos/imunologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Estereoisomerismo , beta-Glucosidase/antagonistas & inibidores
6.
PLoS One ; 14(5): e0217306, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31141557

RESUMO

Endobronchial stents are increasingly used to treat airway complications in multiple conditions including lung transplantation but little is known about the biofilms that form on these devices. We applied deep sequencing to profile luminal biofilms of 46 endobronchial stents removed from 20 subjects primarily with lung transplantation-associated airway compromise. Microbial communities were analyzed by bacterial 16S rRNA and fungal ITS marker gene sequencing. Corynebacterium was the most common bacterial taxa across biofilm communities. Clustering analysis revealed three bacterial biofilm types: one low diversity and dominated by Corynebacterium; another was polymicrobial and characterized by Staphylococcus; and the third was polymicrobial and associated with Pseudomonas, Streptococcus, and Prevotella. Biofilm type was significantly correlated with stent material: covered metal with the Staphylococcus-type biofilm, silicone with the Corynebacterium-dominated biofilm, and uncovered metal with the polymicrobial biofilm. Subjects with sequential stents had frequent transitions between community types. Fungal analysis found Candida was most prevalent, Aspergillus was common and highly enriched in two of three stents associated with airway anastomotic dehiscence, and fungal taxa not typically considered pathogens were highly enriched in some stents. Thus, molecular analysis revealed a complex and dynamic endobronchial stent biofilm with three bacterial types that associate with stent material, a central role for Corynebacterium, and that both expected and unexpected fungi inhabit this unique niche. The current work provides a foundation for studies to investigate the relationship between stent biofilm composition and clinical outcomes, mechanisms of biofilm establishment, and strategies for improved stent technology and use in airway compromise.


Assuntos
Stents/efeitos adversos , Stents/microbiologia , Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Brônquios/microbiologia , Brônquios/cirurgia , Feminino , Fungos/genética , Humanos , Masculino , Microbiota , RNA Ribossômico 16S/genética
7.
Intensive Care Med ; 45(6): 834-843, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31020361

RESUMO

INTRODUCTION: Respiratory tract Candida spp. colonization is associated with more frequent bacterial ventilator-associated pneumonia (VAP). However, this colonization could be causally related to VAP or simply reflect the immune paralysis associated with multiple organ failure. OBJECTIVE: To prospectively evaluate the relationship between Candida spp. colonization and bacterial VAP in mechanically ventilated patients with multiple organ failure. INCLUSION: Patients receiving mechanical ventilation for > 4 days and presenting multiple organ failure were included. Tracheal colonization with Candida spp. was evaluated at inclusion (day 0, D0) and every 4 days until extubation. Quantitative proximal and tracheal cultures were performed at each VAP episode. Monocyte human leukocyte antigen-DR isotype (mHLA-DR) expression and the ratio of polymononuclear leukocytes to lymphocytes were used to evaluate immunoparalysis at D0 and D7. The relationship between fungal colonization and VAP was modelled using cause-specific models for repeated events with adjustment for time-dependent confounders and immune factors. RESULTS: A total of 213 patients, with a median age of 64, simplified acute physiology score II (SAPS II) score 55 and sequential organ failure assessment (SOFA) score 10, mainly admitted for medical reasons (n = 197, 92%), were enrolled in 2012-2015. The median ICU stay was 24 days and the mortality rate was 32% (69 cases). Median mHLA-DR was 5916 Ab-bound/cell [3863-8934]; median lymphocyte count, 0.9Giga/L [0.6-1.3]; neutrophil-to-lymphocyte ratio, 10.9 [6.5-19.7]. Overall, 146 cases (68.5%) had tracheal colonization with Candida spp. An episode of VAP occurred (either for the first or only time) in 62 (29.1%) cases 5.5 days (median) after D0; a second episode occurred in 12 (5.6%) cases, 15.5 days (median) after D0. After adjustment, bronchial colonization with Candida was not associated with VAP [adjusted cause-specific hazard ratio = 0.98 (0.59-1.65), p = 0.95]. CONCLUSION: In patients with mechanical ventilation for more than 4 days and multiple organ failure, bronchial colonization with Candida spp. was not associated with VAP, even after adjustment for immune function.


Assuntos
Candidíase/complicações , Pneumonia Associada à Ventilação Mecânica/etiologia , Adulto , Brônquios/efeitos dos fármacos , Brônquios/microbiologia , Candida/efeitos dos fármacos , Candida/patogenicidade , Candidíase/epidemiologia , Candidíase/fisiopatologia , Estudos de Coortes , Feminino , França/epidemiologia , Humanos , Unidades de Terapia Intensiva/organização & administração , Unidades de Terapia Intensiva/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Escores de Disfunção Orgânica , Pneumonia Associada à Ventilação Mecânica/epidemiologia , Pneumonia Associada à Ventilação Mecânica/fisiopatologia , Estudos Prospectivos , Fatores de Risco , Escala Psicológica Aguda Simplificada
8.
Vet Microbiol ; 232: 50-57, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31030844

RESUMO

Mycoplasma (M.) hyopneumoniae is the initiator agent of the porcine respiratory disease complex (PRDC) and the etiological agent of enzootic pneumonia. M. hyorhinis and M. flocculare are also found in extensive gross pneumonia-like lesions, but their role is not known. We investigated the pathogenicity of M. hyorhinis and M. flocculare in specific-pathogen-free pigs pre-infected or not with M. hyopneumoniae. Mono-inoculated pigs with M. flocculare showed no clinical signs, hematological changes or macroscopic lesions upon necropsy. Mono-inoculated pigs with M. hyorhinis showed, overall seven days after inoculation, an increase in mean temperature with increases in white blood cell (monocyte) counts and in concentrations of pig major acute phase protein, whereas the average daily weight gain (ADWG) decreased compared with non-infected animals. M. hyorhinis was detected in serous membranes (polyserositis) but not in bronchi. Co-infected pigs with M. hyopneumoniae and M. hyorhinis or M. flocculare showed lower ADWG during the third week of the experiment and higher haptoglobin concentrations in contrast to pigs only mono-infected with M. hyopneumoniae. In pigs co-infected with M. hyopneumoniae and M. hyorhinis, it was interesting to observe that (i) M. hyorhinis was detected in bronchi of six pigs, (ii) M. hyopneumoniae was detected in polyserositis and (iii) there was a slight delay in the production of anti-M. hyopneumoniae IgG. The extent of pneumonia was not statistically different between groups. These results suggest that mycoplasmal associations appear to induce an additive effect and increase the inflammatory status in pigs, probably involving in the impairment of the immune system.


Assuntos
Coinfecção/veterinária , Mycoplasma hyopneumoniae/imunologia , Mycoplasma hyorhinis/patogenicidade , Mycoplasma/patogenicidade , Pneumonia Suína Micoplasmática/imunologia , Animais , Anticorpos Antibacterianos/sangue , Brônquios/microbiologia , Coinfecção/microbiologia , Ensaio de Imunoadsorção Enzimática , Haptoglobinas , Pneumonia Suína Micoplasmática/patologia , Organismos Livres de Patógenos Específicos , Suínos , Virulência , Ganho de Peso
9.
Indian J Pathol Microbiol ; 62(2): 326-328, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30971569

RESUMO

Actinomyces naeslundii is a commensal flora of the oral cavity and is generally considered as an avirulent saprophytic bacterium in immunocompetent patients. It can become an opportunistic anaerobic pathogen in oral cavity in patients with poor oral hygiene or tooth extraction and can cause periodontal disease. Pulmonary Actinomycosis is a rare manifestation and may be suspected in middle-aged male patients with cough and hemoptysis showing radiological findings of a peripheral mass or chronic consolidation in whom repeated aerobic cultures have yielded negative results. Here, we report isolation of A. naeslundii from the bronchoalveolar lavage sample from an immunocompetent patient who presented with chronic nonresolving pneumonia of 6 months duration.


Assuntos
Actinomicose/complicações , Actinomicose/diagnóstico , Brônquios/microbiologia , Pneumonia/microbiologia , Actinomyces/isolamento & purificação , Actinomyces/patogenicidade , Brônquios/patologia , Líquido da Lavagem Broncoalveolar/microbiologia , Feminino , Humanos , Imunocompetência , Pulmão/diagnóstico por imagem , Pulmão/microbiologia , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X
10.
Infect Immun ; 87(6)2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30962401

RESUMO

The Gram-negative bacterium Mannheimia haemolytica is the primary bacterial species associated with bovine respiratory disease (BRD) and is responsible for significant economic losses to livestock industries worldwide. Healthy cattle are frequently colonized by commensal serotype A2 strains, but disease is usually caused by pathogenic strains of serotype A1. For reasons that are poorly understood, a transition occurs within the respiratory tract and a sudden explosive proliferation of serotype A1 bacteria leads to the onset of pneumonic disease. Very little is known about the interactions of M. haemolytica with airway epithelial cells of the respiratory mucosa which might explain the different abilities of serotype A1 and A2 strains to cause disease. In the present study, host-pathogen interactions in the bovine respiratory tract were mimicked using a novel differentiated bovine bronchial epithelial cell (BBEC) infection model. In this model, differentiated BBECs were inoculated with serotype A1 or A2 strains of M. haemolytica and the course of infection followed over a 5-day period by microscopic assessment and measurement of key proinflammatory mediators. We have demonstrated that serotype A1, but not A2, M. haemolytica invades differentiated BBECs by transcytosis and subsequently undergoes rapid intracellular replication before spreading to adjacent cells and causing extensive cellular damage. Our findings suggest that the explosive proliferation of serotype A1 M. haemolytica that occurs within the bovine respiratory tract prior to the onset of pneumonic disease is potentially due to bacterial invasion of, and rapid proliferation within, the mucosal epithelium. The discovery of this previously unrecognized mechanism of pathogenesis is important because it will allow the serotype A1-specific virulence determinants responsible for invasion to be identified and thereby provide opportunities for the development of new strategies for combatting BRD aimed at preventing early colonization and infection of the bovine respiratory tract.


Assuntos
Células Epiteliais/microbiologia , Mannheimia haemolytica/patogenicidade , Pasteurelose Pneumônica/microbiologia , Animais , Brônquios/citologia , Brônquios/microbiologia , Bovinos , Mannheimia haemolytica/crescimento & desenvolvimento , Mannheimia haemolytica/fisiologia , Sistema Respiratório/microbiologia , Virulência
11.
mSphere ; 4(1)2019 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728282

RESUMO

Aspergillus fumigatus is a ubiquitous mold that produces small airborne conidia capable of traversing deep into the respiratory system. Recognition, processing, and clearance of A. fumigatus conidia by bronchial airway epithelial cells are thought to be relevant to host defense and immune signaling. Using z-stack confocal microscopy, we observed that only 10 to 20% of adherent conidia from the AF293 clinical isolate are internalized by BEAS-2B cells 6 h postchallenge and not prior. Similar percentages of internalization were observed for the CEA10 clinical isolate. A large subset of both AF293 and CEA10 conidia are rendered metabolically inactive without internalization at 3 h postchallenge by BEAS-2B cells. A significantly larger percentage of CEA10 conidia are metabolically active at 9 and 12 h postchallenge in comparison to the AF293 isolate, demonstrating heterogeneity among clinical isolates. We identified 7 host markers (caveolin, flotillin-2, RAB5C, RAB8B, RAB7A, 2xFYVE, and FAPP1) that consistently localized around internalized conidia 9 h postchallenge. Transient gene silencing of RAB5C, PIK3C3, and flotillin-2 resulted in a larger population of metabolically active conidia. Our findings emphasize the abundance of both host phosphatidylinositol 3-phosphate (PI3P) and PI4P around internalized conidia, as well as the importance of class III PI3P kinase for conidial processing. Therapeutic development focused on RAB5C-, PIK3C3-, and flotillin-2-mediated pathways may provide novel opportunities to modulate conidial processing and internalization. Determination of how contacted, external conidia are processed by airway epithelial cells may also provide a novel avenue to generate host-targeted therapeutics.IMPORTANCE Conidia from the fungus Aspergillus fumigatus are notorious for their ability to stay airborne. This characteristic is believed to allow conidia to penetrate into the cleanest environments. Several hundred conidia are thought to be inhaled each day by a given individual and then expelled by mucociliary clearance. Given that airway epithelial cells make up a significant portion of the pulmonary-air interface, we set out to determine the percentage of conidia that are actually internalized after initial contact with airway epithelial cells. We determined this through an in vitro assay using an immortalized bronchial airway epithelial cell line known as BEAS-2B. Our results suggest a small fraction of conidia are internalized by BEAS-2B cells, while the majority stay adherent to the surface of cells or are washed away during sample processing. Internalization of conidia was observed at 6 h postchallenge and not prior. Our data also indicate conidia are rendered metabolically inactive within 3 h of challenge, suggesting BEAS-2B cells process a large number of conidia without internalization in this early time frame. We have also identified several host endocytosis markers that localize around internalized conidia as well as contribute to the processing of conidia. Understanding how these host endocytosis markers affect the processing of internal and/or external conidia may provide a novel avenue for therapeutic development.


Assuntos
Aspergillus fumigatus/patogenicidade , Endocitose , Células Epiteliais/microbiologia , Animais , Biomarcadores , Brônquios/citologia , Brônquios/microbiologia , Caveolina 1/genética , Linhagem Celular , Células Cultivadas , Interações Hospedeiro-Patógeno , Humanos , Macrófagos/microbiologia , Masculino , Proteínas de Membrana/genética , Camundongos , Fosfatos de Fosfatidilinositol/genética , Aspergilose Pulmonar/microbiologia , Esporos Fúngicos/patogenicidade
12.
PLoS One ; 14(1): e0211290, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30682135

RESUMO

Tobramycin is commonly used to treat Pseudomonas aeruginosa lung infections in patients with Cystic Fibrosis (CF). Tobramycin treatment leads to increased lung function and fewer clinical exacerbations in CF patients, and modestly reduces the density of P. aeruginosa in the lungs. P. aeruginosa resides primarily in the mucus overlying lung epithelial cells and secretes outer membrane vesicles (OMVs) that diffuse through the mucus and fuse with airway epithelial cells, thus delivering virulence factors into the cytoplasm that modify the innate immune response. The goal of this study was to test the hypothesis that Tobramycin reduces the abundance of virulence factors in OMVs secreted by P. aeruginosa. Characterization of the proteome of OMVs isolated from control or Tobramycin-exposed P. aeruginosa strain PAO1 revealed that Tobramycin reduced several OMV-associated virulence determinants, including AprA, an alkaline protease that enhances P. aeruginosa survival in the lung, and is predicted to contribute to the inhibitory effect of P. aeruginosa on Phe508del-CFTR Cl- secretion by primary human bronchial epithelial cells. Deletion of the gene encoding AprA reduced the inhibitory effect of P. aeruginosa on Phe508del-CFTR Cl- secretion. Moreover, as predicted by our proteomic analysis, OMVs isolated from Tobramycin treated P. aeruginosa had a diminished inhibitory effect on Phe508del-CFTR Cl- secretion compared to OMVs isolated from control P. aeruginosa. Taken together, our proteomic analysis of OMVs and biological validation suggest that Tobramycin may improve lung function in CF patients infected with P. aeruginosa by reducing several key virulence factors in OMVs that reduce CFTR Cl- secretion, which is essential for bacterial clearance from the lungs.


Assuntos
Exopeptidases/metabolismo , Proteômica/métodos , Pseudomonas aeruginosa/patogenicidade , Vesículas Secretórias/microbiologia , Tobramicina/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brônquios/citologia , Brônquios/metabolismo , Brônquios/microbiologia , Células Cultivadas , Fibrose Cística/genética , Fibrose Cística/microbiologia , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Exopeptidases/genética , Humanos , Imunidade Inata/efeitos dos fármacos , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/metabolismo , Vesículas Secretórias/efeitos dos fármacos , Vesículas Secretórias/metabolismo , Virulência/efeitos dos fármacos
13.
Am J Physiol Lung Cell Mol Physiol ; 316(1): L206-L215, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30358440

RESUMO

Pseudomonas aeruginosa secretes outer-membrane vesicles (OMVs) that fuse with cholesterol-rich lipid rafts in the apical membrane of airway epithelial cells and decrease wt-CFTR Cl- secretion. Herein, we tested the hypothesis that a reduction of the cholesterol content of CF human airway epithelial cells by cyclodextrins reduces the inhibitory effect of OMVs on VX-809 (lumacaftor)-stimulated Phe508del CFTR Cl- secretion. Primary CF bronchial epithelial cells and CFBE cells were treated with vehicle, hydroxypropyl-ß-cyclodextrin (HPßCD), or methyl-ß-cyclodextrin (MßCD), and the effects of OMVs secreted by P. aeruginosa on VX-809 stimulated Phe508del CFTR Cl- secretion were measured in Ussing chambers. Neither HPßCD nor MßCD were cytotoxic, and neither altered Phe508del CFTR Cl- secretion. Both cyclodextrins reduced OMV inhibition of VX-809-stimulated Phe508del-CFTR Cl- secretion when added to the apical side of CF monolayers. Both cyclodextrins also reduced the ability of P. aeruginosa to form biofilms and suppressed planktonic growth of P. aeruginosa. Our data suggest that HPßCD, which is in clinical trials for Niemann-Pick Type C disease, and MßCD, which has been approved by the U.S. Food and Drug Administration for use in solubilizing lipophilic drugs, may enhance the clinical efficacy of VX-809 in CF patients when added to the apical side of airway epithelial cells, and reduce planktonic growth and biofilm formation by P. aeruginosa. Both effects would be beneficial to CF patients.


Assuntos
2-Hidroxipropil-beta-Ciclodextrina/farmacologia , Micropartículas Derivadas de Células , Cloretos/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Fibrose Cística , Microdomínios da Membrana , Pseudomonas aeruginosa/fisiologia , beta-Ciclodextrinas/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Brônquios/metabolismo , Brônquios/microbiologia , Brônquios/patologia , Linhagem Celular , Micropartículas Derivadas de Células/metabolismo , Micropartículas Derivadas de Células/patologia , Fibrose Cística/tratamento farmacológico , Fibrose Cística/metabolismo , Fibrose Cística/microbiologia , Fibrose Cística/patologia , Células Epiteliais/metabolismo , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Humanos , Microdomínios da Membrana/metabolismo , Microdomínios da Membrana/patologia
14.
Laryngoscope ; 129(3): 544-550, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30284279

RESUMO

OBJECTIVES/HYPOTHESIS: To determine the relationship between mucosal-associated sinus and bronchial microbiota in cystic fibrosis (CF) patients compared to non-CF patients with chronic rhinosinusitis (CRS). STUDY DESIGN: Case series. METHODS: We examined the microbial composition of 52 paired sinus and bronchial brushings from 26 patients with CRS. Paired airway samples from nine subjects with CF were compared with samples from 17 non-CF-CRS disease control patients. The Illumina MiSeq platform was used to sequence the V4 region of the 16S rRNA gene. Sequences were analyzed using QIIME 1.9.0. RESULTS: CF patients demonstrate increased severity of sinus inflammation (Lund-Mackay score 16.3 vs. 12.4, P = .023) and diminished diversity of microbial communities in both the sinuses (Shannon diversity 0.98 vs. 3.3, P = .009) and lungs (Shannon diversity 2.2 vs. 4.0, P = .042) relative to non-CF-CRS. Non-CF-CRS sinus and lung microbiota were distinct and clustered by niche (sinus vs. lung, P = .004). However, CF airway microbiota demonstrated a loss of niche specificity (sinus vs. lung, P = .492). Two CF patients underwent lung transplantation at 4.5 and 9 months prior to sampling. Sinus and lung samples from these two patients demonstrated distinct microbial communities. CONCLUSIONS: Patients with CF undergoing surgery for CRS exhibit substantial bacterial community collapse in the sinuses and a loss of niche specificity between the upper and lower airways compared to non-CF patients with CRS. These results extend previous studies elucidating the lower airway microbiome in cystic fibrosis and provide support for the concept of microbial translocation in the cystic fibrosis airways. LEVEL OF EVIDENCE: 4 Laryngoscope, 129:544-550, 2019.


Assuntos
Bactérias/isolamento & purificação , Brônquios/microbiologia , Fibrose Cística/microbiologia , Microbiota , Seios Paranasais/microbiologia , Rinite/microbiologia , Sinusite/microbiologia , Doença Crônica , Fibrose Cística/complicações , Humanos , Estudos Prospectivos , Rinite/complicações , Sinusite/complicações
15.
Rev Mal Respir ; 36(1): 63-68, 2019 Jan.
Artigo em Francês | MEDLINE | ID: mdl-30429094

RESUMO

INTRODUCTION: Bronchial mould infection or plastic bronchitis is a rare condition, encountered at any age, but with a predilection for childhood. The clinical diagnosis is made easy by the demonstration of solid, branched expectorations. However, the aetiology is not easy to determine and investigation does not often lead to a pathological diagnosis. CASE REPORT: We report the case of a 24 year-old patient, with a history of pulmonary tuberculosis in January 2016, who had had chronic, persistent, solid and branched expectorations since January 2015. Fibreoptic bronchoscopy revealed thick white secretions plugging the bronchi. Pathological examination of the bronchial plugs showed fibrous tissue infiltrated with predominantly lymphoplasmocytic and histiocytic inflammatory cells. Investigations carried out on the pleura did not establish the aetiological diagnosis. The diagnosis of bronchial mould disease of tuberculous origin complicated by pleurisy was established. Corticosteroid therapy led to a complete regression of the moulds within six weeks. CONCLUSION: The presence of solid and branched sputum should suggest fungal bronchitis and is an indication for bronchoscopy for diagnostic and therapeutic purposes. Early management is important to avoid complications.


Assuntos
Brônquios/patologia , Bronquite/diagnóstico , Empiema Pleural/etiologia , Tuberculose Pulmonar/complicações , Adulto , Antibacterianos/uso terapêutico , Brônquios/microbiologia , Bronquite/tratamento farmacológico , Bronquite/etiologia , Broncoscopia/métodos , Diagnóstico Diferencial , Empiema Pleural/tratamento farmacológico , Glucocorticoides/uso terapêutico , Humanos , Masculino , Escarro/microbiologia , Tomografia Computadorizada por Raios X , Tuberculose Pulmonar/tratamento farmacológico
16.
J Mycol Med ; 29(2): 112-119, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30446390

RESUMO

To study invasive pulmonary Aspergillosis (IPA), we depleted neutrophils in mice using the monoclonal antibody anti-Gr-1/Ly-6G. Immunocompetent and neutropenic mice were infected via intratracheal with conidia of Aspergillus fumigatus clinical isolates, characterized as either higher or lower elastase producers. Neutropenic animals exhibited 100% mortality in 5 days, for both strains, and were observed survival curves overlapped, lungs with angioinvasion, rupture of bronchial and vascular walls, associated with exuberance of conidia filamentation. The immunocompetent animals infected with the lower elastase producer strain presented with upregulated inflammatory processes, and a lack of conidia filamentation in the tissue. The fungal burden in the lungs was not different in the immunocompetent and neutropenic groups. These findings confirm the protective role of neutrophils against A. fumigatus and suggest that the fungal elastinolytic activity is not a critical virulence factor but may be involved in tissue injury.


Assuntos
Aspergillus fumigatus/isolamento & purificação , Modelos Animais de Doenças , Aspergilose Pulmonar Invasiva/mortalidade , Aspergilose Pulmonar Invasiva/patologia , Camundongos , Animais , Aspergillus fumigatus/enzimologia , Brônquios/microbiologia , Brônquios/patologia , Humanos , Aspergilose Pulmonar Invasiva/microbiologia , Pulmão/microbiologia , Pulmão/patologia , Masculino , Camundongos Endogâmicos C57BL , Neutropenia , Neutrófilos/imunologia , Elastase Pancreática/metabolismo , Esporos Fúngicos/isolamento & purificação , Esporos Fúngicos/fisiologia
17.
J Med Microbiol ; 68(2): 241-247, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30526740

RESUMO

PURPOSE: Pertussis remains a public health concern in most countries. Our study aimed to prospectively explore the epidemiology of pertussis in the Tunis area of Tunisia between 2007 and 2016, and to characterize the virulence-associated genes of the collected Bordetella pertussis isolates. METHODOLOGY: Infants and children hospitalized at the Children's Hospital of Tunis, Tunisia, between 2007 and 2016 for suspicion of pertussis were enrolled in the study. Culture and real-time PCR (qPCR) assays targeting IS481, IS1001, recA, H-IS1001 and ptxP were used to confirm the pertussis diagnosis. Phenotypic and genotypic characterization of recovered isolates was performed.Results/Key findings. A total of 1844 children were included in the study. Overall, 306 children (16.6 %) with Bordetella infection were confirmed by qPCR. Among them, 265 (86.6 %) were confirmed as having B. pertussis (IS481+, ptxP+, H-IS1001-), 18 (5.9 %) as having Bordetella parapertussis (IS481-, IS1001+) and 11 (3.6 %) as having Bordetella spp. (IS481+, ptxP-, H-IS1001-). No Bordetella holmesii (IS481+, IS1001-, H-IS1001+) was identified. The estimated pertussis incidence in the Tunis area was 134/100 000 in children aged less than 5 years. Two epidemic peaks were observed in 2009 and 2014. Ten B. pertussis isolates were cultured and characterized. Deficiency in pertactin expression was not observed, and genotyping of the isolates revealed a predominant allelic profile: ptxP3-ptxA1-prn2-fim2-1-fim3-2. CONCLUSION: This study demonstrated that pertussis is still present as a cyclical disease in Tunisia, despite high primo-vaccination coverage with a pertussis whole-cell vaccine. The predominant genotype of Tunisian B. pertussis isolates is similar to isolates circulating in countries using the acellular vaccine.


Assuntos
Bordetella pertussis/isolamento & purificação , Coqueluche/epidemiologia , Coqueluche/microbiologia , Antibacterianos/farmacologia , Bordetella pertussis/classificação , Bordetella pertussis/efeitos dos fármacos , Brônquios/microbiologia , Criança , Pré-Escolar , Tosse , Cianose , Feminino , Genótipo , Humanos , Immunoblotting , Lactente , Recém-Nascido , Masculino , Nasofaringe/microbiologia , Fenótipo , Reação em Cadeia da Polimerase , Estudos Prospectivos , Traqueia/microbiologia , Tunísia/epidemiologia
19.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30573530

RESUMO

Mycoplasma pneumoniae is one of the most common pathogenic causes of community-acquired pneumonia. Hydrogen sulfide, alanine, and pyruvate producing enzyme (HapE) is a recently discovered M. pneumoniae virulence factor that can produce H2S to promote erythrocyte lysis. However, other cytotoxic effects of HapE have not been explored. The present study examined the effects of this enzyme on normal human bronchial epithelial (NHBE) cells, in an attempt to identify additional mechanisms of M. pneumoniae pathogenesis. Recombinant HapE was purified for use in downstream assays. MTT and colony formation assays were conducted to determine the effects of HapE on cell viability and growth, while flow cytometry was used to examine changes in cell proliferation and cell cycle function. ELISA was performed to examine changes in the cytokine profile of HapE-treated cells. HapE treatment arrested NHBE cells in S phase and inhibited cell proliferation in a concentration-dependent manner. The anti-inflammatory factors interleukin (IL)-4 and IL-6 were significantly enhanced following HapE treatment. Increased secretion of pro-inflammatory factors was not observed. The effects of HapE on the respiratory epithelium may have an impact on the efficiency of host immune surveillance and pathogen elimination, and contribute to the pathogenesis of M. pneumoniae.


Assuntos
Proteínas de Bactérias/genética , Brônquios/microbiologia , Mycoplasma pneumoniae/enzimologia , Pneumonia por Mycoplasma/enzimologia , Fatores de Virulência/genética , Proteínas de Bactérias/farmacologia , Brônquios/enzimologia , Proliferação de Células/genética , Citocinas/genética , Citocinas/farmacologia , Células Epiteliais/enzimologia , Células Epiteliais/microbiologia , Humanos , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/patogenicidade , Pneumonia por Mycoplasma/genética , Pneumonia por Mycoplasma/microbiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Fatores de Virulência/farmacologia
20.
PLoS One ; 13(11): e0208277, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30500862

RESUMO

Mouse models have been essential to generate supporting data for the research of infectious diseases. Burkholderia pseudomallei, the etiological agent of melioidosis, has been studied using mouse models to investigate pathogenesis and efficacy of novel medical countermeasures to include both vaccines and therapeutics. Previous characterization of mouse models of melioidosis have demonstrated that BALB/c mice present with an acute infection, whereas C57BL/6 mice have shown a tendency to be more resistant to infection and may model chronic disease. In this study, either BALB/c or C57BL/6 mice were exposed to aerosolized human clinical isolates of B. pseudomallei. The bacterial strains included HBPUB10134a (virulent isolate from Thailand), MSHR5855 (virulent isolate from Australia), and 1106a (relatively attenuated isolate from Thailand). The LD50 values were calculated and serial sample collections were performed in order to examine the bacterial burdens in tissues, histopathological features of disease, and the immune response mounted by the mice after exposure to aerosolized B. pseudomallei. These data will be important when utilizing these models for testing novel medical countermeasures. Additionally, by comparing highly virulent strains with attenuated isolates, we hope to better understand the complex disease pathogenesis associated with this bacterium.


Assuntos
Burkholderia pseudomallei/fisiologia , Melioidose/patologia , Animais , Formação de Anticorpos , Austrália/epidemiologia , Brônquios/imunologia , Brônquios/microbiologia , Brônquios/patologia , Burkholderia pseudomallei/patogenicidade , Citocinas/sangue , Citocinas/imunologia , Modelos Animais de Doenças , Progressão da Doença , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Melioidose/sangue , Melioidose/epidemiologia , Melioidose/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Tailândia/epidemiologia , Virulência
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