Mário Andrea

Contact endoscopy during microlaryngeal surgery : a new technique for endoscopic examination of the larynx

By means of a microcolpohysteroscope during microlaryngoscopy, the cells of superficial layers of the vocal cord epithelium, previously stained with methylene blue, were observed in vivo and in situ. Several parameters were evaluated: regularity of the epithelium, dimensions and color of the nucleus, nucleus-cytoplasm ratio, nucleus and cytoplasm contours, and presence of the nucleolus, mitosis, cytoplasmic inclusions, keratosis, and koilocytes, among others. The normal squamous epithelium of the vocal cord has a homogeneous cell arrangement, with a spheroid and isochromic nucleus, and a uniform nucleus-cytoplasm ratio. Specific cellular epithelial patterns in different disorders were observed, namely, chronic laryngitis, keratosis, dysplasia, papilloma, and malignant tumor. The present study on contact endoscopy during microlaryngeal surgery reports our first results with this original technique, performed in 68 patients.

Contact Endoscopy of the Vocal Cord: Normal and Pathological Patterns

Contact endoscopy allows an in vivo and in situ study of the superficial layers of the vocal cord epithelium. This technique offers access to some characteristics of the epithelium cells: dimension and shape of the nuclei, nucleus/cytoplasm ratio, keratosis, presence of nucleolus, mitosis and koylocites. Contact endoscopy was performed during microlaryngoscopies in 92 patients with benign, pre-malign and malign pathology of the vocal cords. Systematic confrontation with pathological examination and laryngectomy specimens studied by histology and electron microscopy were used to improve the interpretation of contact endoscopy findings. Based on this correlation it has been possible to analyse the transition of the squamous and ciliated epithelium of the vocal cord as well as to delineate some pathological patterns, e.g. chronic laryngitis, keratosis, dysplasia, tumor and papilloma.

Jitter estimation algorithms for detection of pathological voices

This work is focused on the evaluation of different methods to estimate the amount of jitter present in speech signals. The jitter value is a measure of the irregularity of a quasiperiodic signal and is a good indicator of the presence of pathologies in the larynx such as vocal fold nodules or a vocal fold polyp. Given the irregular nature of the speech signal, each jitter estimation algorithm relies on its own model making a direct comparison of the results very difficult. For this reason, the evaluation of the different jitter estimation methods was target on their ability to detect pathological voices. Two databases were used for this evaluation: a subset of the MEEI database and a smaller database acquired in the scope of this work. The results showed that there were significant differences in the performance of the algorithms being evaluated. Surprisingly, in the largest database the best results were not achieved with the commonly used relative jitter, measured as a percentage of the glottal cycle, but with absolute jitter values measured in microseconds. Also, the new proposed measure for jitter, LocJitt, performed in general is equal to or better than the commonly used tools of MDVP and Praat.

Contact Endoscopy of the Nasal Mucosa

The possibility of contact endoscopy in the nasal mucosa, using a contact microlaryngoscope is examined. With contact endoscopy it has been possible to visualize in vivo and in situ (60x, 150x) the superficial cell layers of the nasal epithelium, previously stained with methylene blue. Normal mucosa and cases with pathology (chronic rhinitis, nasal polyposis, inverted papilloma, mucormicosis and carcinoma) were assessed with contact endoscopy (50 patients). Squamous epithelium, ciliated epithelium, glandular ostia, mucus secretions, microvascular networks, inflammatory cell infiltrates, tissue inclusions, nuclear abnormalities, and fungal hyphae have been made visible. The potential of contact endoscopy in the nose, the alterations required in the contact endoscope and the intense interdisciplinary work needed are discussed. Evaluation and differentiation of stages of chronic mucosal diseases, nasal polyposis, environmental pathology, allergy, mucocilliary diseases, pharmacotherapy, are some of the entities that will benefit from this technique.

Endoscopic anatomy of the larynx

Rigid videoendoscopy performed during microlaryngoscopy allows exploration of regions that are difficult to observe. Higher magnifications improve the definition of lesion characteristics. Epithelial cells, which have been previously stained with methylene blue, can be identified with contact endoscopy during microlaryngoscopy. In this new phase of laryngeal endoscopic development, it is possible to perform in vivo and in situ studies of the cells, nucleus, cytoplasm, and microvascular networks.

Proposal for a descriptive guideline of vascular changes in lesions of the vocal folds by the committee on endoscopic laryngeal imaging of the European Laryngological Society

In the last decades new endoscopic tools have been developed to improve the diagnostic work-up of vocal fold lesions in addition to normal laryngoscopy, i.e., contact endoscopy, autofluorescence, narrow band imaging and others. Better contrasted and high definition images offer more details of the epithelial and superficial vascular structure of the vocal folds. Following these developments, particular vascular patterns come into focus during laryngoscopy. The present work aims at a systematic pathogenic description of superficial vascular changes of the vocal folds. Additionally, new nomenclature on vascular lesions of the vocal folds will be presented to harmonize the different terms in the literature. Superficial vascular changes can be divided into longitudinal and perpendicular. Unlike longitudinal vascular lesions, e.g., ectasia, meander and change of direction, perpendicular vascular lesions are characterized by different types of vascular loops. They are primarily observed in recurrent respiratory papillomatosis, and in pre-cancerous and cancerous lesions of the vocal folds. These vascular characteristics play a significant role in the differential diagnosis. Among different parameters, e.g., epithelial changes, increase of volume, stiffness of the vocal fold, vascular lesions play an increasing role in the diagnosis of pre- and cancerous lesions.

A7445G mtDNA mutation present in a Portuguese family exhibiting hereditary deafness and palmoplantar keratoderma

Mitochondrial DNA (mtDNA) A7445G point mutation has been shown to be responsible for familial nonepidermolytic palmoplantar keratoderma (NEPPK) associated with deafness without any additional features. To date, only a few cases have been described. We report a Portuguese pedigree presenting an inherited combination of NEPPK and sensorineural deafness compatible with maternal transmission. Clinical expression and age of onset of NEPPK and deafness were variable. Normal expression patterns of epidermal keratins and filaggrin, intercellular junction proteins including connexin 26, loricrin and cornified envelope proteins, were observed. Molecular analysis revealed that all the affected members, previously screened for Cx26 mutations with negative results, presented the mtDNA A7445G point mutation in the homoplasmic form. To our knowledge, this is the fifth family in whom inherited NEPPK and hearing loss are related to this mitochondrial mutation.

Spectrum and frequency of GJB2 mutations in a cohort of 264 Portuguese nonsyndromic sensorineural hearing loss patients

Abstract Objective: To assess the spectrum and prevalence of mutations in the GJB2 gene in Portuguese nonsyndromic sensorineural hearing loss (NSSHL) patients. Design: Sequencing of the coding region, basal promoter, exon 1, and donor splice site of the GJB2 gene; screening for the presence of the two common GJB6 deletions. Study sample: A cohort of 264 Portuguese NSSHL patients. Results: At least one out of 21 different GJB2 variants was identified in 80 (30.2%) of the 264 patients analysed. Two mutant alleles were found in 53 (20%) of these probands, of which 83% (44/53) harboured at least one c.35delG allele. Twenty-seven (10.2%) of the probands harboured only one mutant allele. Subsequent analysis revealed that the GJB6 deletion del(GJB6-D13S1854) was present in at least 7.4% (2/27) of the patients carrying only one mutant GJB2 allele. Overall, one in five (55/264) of the patients were diagnosed as having DFNB1-related NSSHL, of which the vast majority (53/55) harboured only GJB2 mutations. Conclusions: This study provides clear demonstration that mutations in the GJB2 gene are an important cause of NSSHL in Portugal, thus representing a valuable indicator as regards therapeutical and rehabilitation options, as well as genetic counseling of these patients and their families.

Is the Sagittal Postural Alignment Different in Normal and Dysphonic Adult Speakers

OBJECTIVE Clinical research in the field of voice disorders, in particular functional dysphonia, has suggested abnormal laryngeal posture due to muscle adaptive changes, although specific evidence regarding body posture has been lacking. The aim of our study was to verify if there were significant differences in sagittal spine alignment between normal (41 subjects) and dysphonic speakers (33 subjects). STUDY DESIGN Cross-sectional study. METHODS Seventy-four adults, 35 males and 39 females, were submitted to sagittal plane photographs so that spine alignment could be analyzed through the Digimizer-MedCalc Software Ltd program. Perceptual and acoustic evaluation and nasoendoscopy were used for dysphonic judgments: normal and dysphonic speakers. RESULTS For thoracic length curvature (TL) and for the kyphosis index (KI), a significant effect of dysphonia was observed with mean TL and KI significantly higher for the dysphonic speakers than for the normal speakers. Concerning the TL variable, a significant effect of sex was found, in which the mean of the TL was higher for males than females. The interaction between dysphonia and sex did not have a significant effect on TL and KI variables. For the lumbar length curvature variable, a significant main effect of sex was demonstrated; there was no significant main effect of dysphonia or significant sex×dysphonia interaction. CONCLUSIONS Findings indicated significant differences in some sagittal spine posture measures between normal and dysphonic speakers. Postural measures can add useful information to voice assessment protocols and should be taken into account when considering particular treatment strategies.

Novel splice-site mutation c.1615-2A>G (IVS14-2A>G) in the SLC26A4 gene causing Pendred syndrome in a consanguineous Portuguese family.

Novel Splice-Site Mutation c.1615-2A>G (IVS14-2A>G) in the SLC26A4 Gene Causing Pendred Syndrome in a Consanguineous Portuguese Family Helena Sim~oes-Teixeira, Tiago D. Matos, Marta Canas Marques, Óscar Dias, M ario Andrea, Eduardo Barreiros, Lu ıs Barreiros, Felipe Moreno, Graça Fialho, Helena Caria, and Ignacio del Castillo* Faculty of Science, Centre for Biodiversity, Functional and Integrative Genomics (BioFIG), University of Lisbon, Lisbon, Portugal Unidad de Gen etica Molecular, Hospital Universitario Ram on y Cajal, IRYCIS, Madrid, Spain ORL Service, Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, EPE, Lisbon, Portugal Endocrinology Department, Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, EPE, Lisbon, Portugal Centro de Investigaci on Biom edica en Red de Enfermedades Raras (CIBERER), Madrid, Spain Higher School of Health, Polytechnic Institute of Set ubal, Set ubal, Portugal