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1.
mSphere ; 4(5)2019 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-31484741

RESUMO

Otitis media (OM) is a cluster of diseases of the middle ear that commonly result from bacterial infection. OM subtypes in which the tympanic membrane is intact (acute otitis media and otitis media with effusion) are presumed to result from pathogen translocation through the eustachian tube. Recent molecular-based studies have suggested that a diverse middle ear microbiome exists in the absence of disease. These have been largely unsupported by culture and feature species that commonly contaminate low-biomass sequencing data. Combining culture-based and molecular techniques, we undertook a detailed investigation of the evidence for bacterial colonization of the healthy middle ear. Middle ear (ME), nasopharynx (NP), and external ear canal (EC) swabs were collected from a total of 25 adult patients undergoing cochlear implant, stapedotomy, or translabyrinthine vestibular schwannoma resection. Diagnostic culture, microscopy, quantitative PCR, and 16S rRNA gene amplicon sequencing were used to assess sample bacterial content. EC and NP microbiota were consistent with previous reports. In contrast, bacterial levels in ME samples were not significantly above those in unused control swabs. Commonly detected taxa were among recognized sequencing contaminants (Methylobacterium, Pseudomonas, and Acinetobacter). Linear regression of dominant ME taxa confirmed a negative relationship between relative abundance and bacterial load, consistent with contamination. No bacteria were detected by microscopy or diagnostic culture in any middle ear sample. Our findings cast substantial doubt on previous reports identifying a healthy middle ear microbiome using 16S amplicon sequencing.IMPORTANCE Recent molecular-based studies have suggested that a diverse middle ear microbiome in adults and children can exist in the absence of disease. These studies have been largely unsupported by culture and feature species that commonly contaminate low-biomass sequencing data. While 16S rRNA gene amplicon sequencing has proven to be a highly informative technique in many clinical contexts, it is susceptible to spurious signal arising from sequencing reagent contaminants where sample biomass is low. Combining culture-based and molecular techniques, we undertook a detailed investigation of the evidence for bacterial colonization of the healthy middle ear. In finding no evidence of viable bacterial cells in middle ear samples, our study further underlines the importance of careful consideration of amplicon sequence data derived from very-low-biomass contexts and the value of analytical approaches that combine culture and molecular techniques.


Assuntos
Orelha Média/microbiologia , Microbiota , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias/genética , Bactérias/isolamento & purificação , Orelha Externa/microbiologia , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/microbiologia , RNA Ribossômico 16S/genética , Células-Tronco , Adulto Jovem
2.
Sci Rep ; 6: 34081, 2016 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-27665737

RESUMO

Squamous cell carcinoma (SCC) is the second most common cancer worldwide and accounts for approximately 30% of all keratinocyte cancers. The vast majority of cutaneous SCCs of the head and neck (cSCCHN) are readily curable with surgery and/or radiotherapy unless high-risk features are present. Perineural invasion (PNI) is recognized as one of these high-risk features. The molecular changes during clinical PNI in cSCCHN have not been previously investigated. In this study, we assessed the global gene expression differences between cSCCHN with or without incidental or clinical PNI. The results of the analysis showed signatures of gene expression representative of activation of p53 in tumors with PNI compared to tumors without, amongst other alterations. Immunohistochemical staining of p53 showed cSCCHN with clinical PNI to be more likely to exhibit a diffuse over-expression pattern, with no tumors showing normal p53 staining. DNA sequencing of cSCCHN samples with clinical PNI showed no difference in mutation number or position with samples without PNI, however a significant difference was observed in regulators of p53 degradation, stability and activity. Our results therefore suggest that cSCCHN with clinical PNI may be more likely to contain alterations in the p53 pathway, compared to cSCCHN without PNI.

3.
Cancer Immunol Immunother ; 65(2): 213-22, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26759008

RESUMO

Spread of head and neck cancer along the cranial nerves is often a lethal complication of this tumour. Current treatment options include surgical resection and/or radiotherapy, but recurrence is a frequent event suggesting that our understanding of this tumour and its microenvironment is incomplete. In this study, we have analysed the nature of the perineural tumour microenvironment by immunohistochemistry with particular focus on immune cells and molecules, which might impair anti-tumour immunity. Moderate to marked lymphocyte infiltrates were present in 58.8% of the patient cohort including T cells, B cells and FoxP3-expressing T cells. While human leukocyte antigen (HLA) class I and more variably HLA class II were expressed on the tumour cells, this did not associate with patient survival or recurrence. In contrast, galectin-1 staining within lymphocyte areas of the tumour was significantly associated with a poorer patient outcome. Given the known role of galectin-1 in immune suppression, the data suggest that galectin inhibitors might improve the prognosis of patients with perineural spread of cancer.


Assuntos
Galectina 1/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/mortalidade , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/mortalidade , Idoso , Nervos Cranianos/patologia , Feminino , Neoplasias de Cabeça e Pescoço/imunologia , Neoplasias de Cabeça e Pescoço/patologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Antígenos de Histocompatibilidade Classe II/metabolismo , Humanos , Estimativa de Kaplan-Meier , Subpopulações de Linfócitos/imunologia , Subpopulações de Linfócitos/metabolismo , Subpopulações de Linfócitos/patologia , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/metabolismo , Linfócitos do Interstício Tumoral/patologia , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Invasividade Neoplásica , Recidiva Local de Neoplasia , Avaliação de Resultados da Assistência ao Paciente , Prognóstico , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Microambiente Tumoral
4.
Otolaryngol Head Neck Surg ; 141(2): 257-263, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19643262

RESUMO

OBJECTIVE: To measure long-term quality-of-life (QOL) improvement following contemporary multilevel upper airway reconstruction surgery, compared with continuous positive airway pressure (CPAP) therapy. Secondary aims were to investigate factors determining clinical effectiveness and QOL impact of reported side effects. DESIGN: Cohort study. SUBJECTS AND METHODS: Consecutive, simultaneously treated adult patients with moderate-severe obstructive sleep apnea (OSA) having upper airway surgery (N=77) or CPAP (N=89) therapy were studied by questionnaire. Glasgow Benefit Inventory (GBI), change in snoring status and Epworth Sleepiness Scale (ESS), subjective CPAP compliance, and side effects in both groups were measured at mean+/-SEM 44.12+/-5.78 months (3.68+/-0.48 years) after commencement of therapy. RESULTS: No significant difference was seen between surgical outcomes for GBI, snoring, or ESS and CPAP controls. Multivariate analysis showed reduction in Respiratory Disturbance Index (RDI) predicted postoperative snoring and ESS, but not GBI outcomes. Snoring control and GBI were related to CPAP compliance (P<0.001). CPAP side effects (reported in 26%) significantly reduced the QOL benefit of treatment, independent of compliance. Surgical complications (occurring in 44%) did not affect QOL treatment benefit. CONCLUSION: Patients with poor CPAP compliance and/or significant side effects of CPAP therapy (45% of cases in this series) should be evaluated for contemporary upper airway reconstructive surgery.


Assuntos
Pressão Positiva Contínua nas Vias Aéreas , Procedimentos Cirúrgicos Bucais/métodos , Qualidade de Vida , Apneia Obstrutiva do Sono/cirurgia , Ronco/cirurgia , Estudos de Coortes , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Polissonografia , Procedimentos de Cirurgia Plástica/métodos , Apneia Obstrutiva do Sono/terapia , Ronco/terapia , Inquéritos e Questionários , Resultado do Tratamento
5.
Head Neck Oncol ; 1: 12, 2009 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-19442314

RESUMO

The principles of management of the laryngeal cancer have evolved over the recent past with emphasis on organ preservation. These developments have paralleled technological advancements as well as refinement in the surgical technique. The surgeons are able to maintain physiological functions of larynx namely speech, respiration and swallowing without compromising the loco-regional control of cancer in comparison to the more radical treatment modalities. A large number of organ preservation surgeries are available to the surgeon; however, careful assessment of the stage of the cancer and selection of the patient is paramount to a successful outcome. A comprehensive review of various organ preservation techniques in vogue for the management of laryngeal cancer is presented.


Assuntos
Neoplasias Laríngeas/cirurgia , Preservação de Órgãos , Ablação por Cateter , Terapia Combinada , Humanos , Neoplasias Laríngeas/diagnóstico , Laringectomia/métodos , Laringoscopia , Procedimentos de Cirurgia Plástica/métodos , Robótica
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