The serine protease hepsin mediates urinary secretion and polymerisation of Zona Pellucida domain protein uromodulin.

Uromodulin is the most abundant protein in the urine. It is exclusively produced by renal epithelial cells and it plays key roles in kidney function and disease. Uromodulin mainly exerts its function as an extracellular matrix whose assembly depends on a conserved, specific proteolytic cleavage leading to conformational activation of a Zona Pellucida (ZP) polymerisation domain. Through a comprehensive approach, including extensive characterisation of uromodulin processing in cellular models and in specific knock-out mice, we demonstrate that the membrane-bound serine protease hepsin is the enzyme responsible for the physiological cleavage of uromodulin. Our findings define a key aspect of uromodulin biology and identify the first in vivo substrate of hepsin. The identification of hepsin as the first protease involved in the release of a ZP domain protein is likely relevant for other members of this protein family, including several extracellular proteins, as egg coat proteins and inner ear tectorins.

The effects of noise exposure on nitric oxide synthase knockout mice

This paper studies the role of nitric oxide (NOS 1, NOS 2, and NOS 3 genes) in the mouse cochlea and in noise-induced hearing loss (NIHL). Mice genetically deficient of the NOS 2 and NOS 3 genes were protected from NIHL, indicating that one or both of these genes may be responsible for producing nitric oxide that damages the inner ear when exposed to harmful levels of noise.

Telefones celulares: influência nos sistemas auditivo e vestibular

Os sistemas de telecomunicações emitem radiofreqüência, uma radiação eletromagnética invisível. Telefones celulares transmitem microondas (450900 MHz no sistema analógico e 1,82,2 GHz no sistema digital), muito próximo à orelha do usuário. Esta energia é absorvida pela pele, orelha interna, nervo vestibulococlear e superfície do lobo temporal. OBJETIVO: Revisar a literatura sobre influência dos telefones celulares na audição e equilíbrio. FORMA DE ESTUDO: Revisão sistemática. METODOLOGIA: Foram pesquisados artigos nas bases Lilacs e Medline sobre a influência dos telefones celulares nos sistemas auditivo e vestibular, publicados de 2000 a 2005, e também materiais veiculados na Internet. RESULTADOS: Os estudos sobre radiação do telefone celular e risco de neurinoma do acústico apresentam resultados contraditórios. Alguns autores não encontram maior probabilidade de aparecimento do tumor nos usuários de celulares, enquanto outros relatam que a utilização de telefones analógicos por 10 anos ou mais aumenta o risco para o tumor. A exposição aguda às microondas emitidas pelo celular não influencia a atividade das células ciliadas externas da cóclea, in vivo e in vitro, a condução elétrica no nervo coclear, nem a fisiologia do sistema vestibular em humanos. As próteses auditivas analógicas são mais suscetíveis à interferência eletromagnética dos telefones celulares digitais. CONCLUSÃO: Não há comprovação de lesão cocleovestibular pelos telefones celulares.

Ontogenia do sistema sensorial de pacu Piaractus mesopotamicus (Holmberg, 1887) (Characidae: Serrasalmidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise morfométrica do sistema auditivo periférico da preguiça (Bradypus variegatus)

A superordem Xenarthra é composta de 31 espécies viventes de tatus, tamanduás e preguiças. As arborícolas preguiças pertencem a dois gêneros, Choloepus e Bradypus, cuja divergência se deu a aproximadamente 40 milhões de anos atrás. As similaridades entre os dois taxa, tais como a presença de algas verdes nos pêlos e habilidade locomotora suspensória, são notáveis exemplos de evolução convergente. A exata posição da linhagem Xenarthra entre os mamíferos na árvore filogenética ainda não é completamente compreendida, com alguns rearranjos na árvore da família dos mamíferos placentários, considerando os Xenarthras mais relacionados entre Afrotheria (que inclui musaranhos, porcos-da-terra, peixes-boi e elefantes) ou a Boreoeutheria (que inclui primatas, roedores, carnívoros e ungulados). O objetivo deste trabalho é descrever pela primeira vez características morfológicas dos ouvidos médio e interno de Bradypus variegatus e compará-las a outros mamíferos placentários que possuam dados publicados na literatura. Nós usamos 13 espécimes adultas post-mortem (machos e fêmeas) e 15 crânios da coleção do Museu Paraense Emílio Goeldi. Além das medições, foram usadas técnicas de microscopia óptica, microscopia eletrônica de varredura e tomografia computadorizada. Através da árvore filogenética das preguiças, o gênero Bradypus é posicionado como táxon-irmão de todas as outras preguiças. Nossos resultados mostram que a morfologia do ouvido médio e interno de Bradypus variegatus é similar a de outros mamíferos com dados publicados na literatura e que apresentam escalonamento alométrico.

Anatomia macroscópica e ultraestrutural da orelha da paca (Cuniculus paca, Linnaeus, 1766)

Pós-graduação em Cirurgia Veterinária - FCAV

Produção e utilização de material didático sobre fisiologia da audição e neurofisiologia

The teaching of hearing physiology requires an knowledge integration of Human Anatomy, Biophysics, more precisely Bioacoustics and Bioelectrogenesis, as well as Neurophysiology. Students present difficulty to build knowledge about functional mechanisms of sound conduction and sensory transduction, especially if the elements are not visible forms, as the middle and inner ear structures. To make the teaching about hearing physiology and sensory perception easier, was produced a set of didactical materials about the subject. At first, a resin model that faithfully describes the anatomical relationship of the ossicles with the tympanic membrane was developed. Subsequently, a second model that, besides illustrates the mechanism of acoustic impedance overcoming, also reveals how acoustic sensorial transduction occurs in inner ear, was designed and produced. In the third didactical model, are visualized, through students interaction, areas of the cerebral cortex that interpret the different sensory modalities. In addition, were created three educational videos about hearing problems and a site on Human Hearing Physiology, available on Institute of Biosciences website. The results of this course conclusion monograph are presented in the form of articles that were submitted to Journal Physics in the School and the Journal of the Nucleus of Teaching

Morphological aspects of tympanic bulla after ventral osteotomy in cats

To evaluate tympanic bulla healing after experimental ventral osteotomy in cats. Twenty adult cats were submitted to unilateral ventral bulla osteotomy and divided into two groups: cats of A1 group (n=10) were euthanized at 8 weeks and cats of A2 group (n=10), at 16 weeks postoperative. Signs of Horner's syndrome or damage to the inner ear were not found. Open-mouth radiographs taken in the immediate postoperative showed interruption in the contour of the larger compartment of the operated bulla. The result of Mcnemar'test was significant in A2 group (*p=0.0156). Macroscopic exams revealed that the operated bullae were similar to the normal ones, with preservation of the tympanic cavity. Connective tissue at the osteotomy site of the larger compartment was significantly found in the operated bullae in both groups (McNemar test: A1 p=0.0020*; A2 p=0.0078*). Histomorphometric analyses showed that the connective tissue length at the osteotomy site was shorter in A2 group than in the A1 group (Mann-Whitney test: p=0.0021*). Experimental ventral osteotomy did not alter significantly the tympanic bulla conformation and complete regeneration of tympanic bulla frequently did not occur before 16 weeks of postoperative period.

Design of a semi-implantable hearing device for direct acoustic cochlear stimulation

A new hearing therapy based on direct acoustic cochlear stimulation was developed for the treatment of severe to profound mixed hearing loss. The device efficacy was validated in an initial clinical trial with four patients. This semi-implantable investigational device consists of an externally worn audio processor, a percutaneous connector, and an implantable microactuator. The actuator is placed in the mastoid bone, right behind the external auditory canal. It generates vibrations that are directly coupled to the inner ear fluids and that, therefore, bypass the external and the middle ear. The system is able to provide an equivalent sound pressure level of 125 dB over the frequency range between 125 and 8000 Hz. The hermetically sealed actuator is designed to provide maximal output power by keeping its dimensions small enough to enable implantation. A network model is used to simulate the dynamic characteristics of the actuator to adjust its transfer function to the characteristics of the middle ear. The geometry of the different actuator components is optimized using finite-element modeling.

Molecular analysis of aquaporin genes 1 to 4 in patients with Menière's disease

Menière's Disease (MD) is an episodic cochleovestibular dysfunction of unknown etiology, still lacking a specific test and therapy. The proposed theories on the pathophysiology include genetic factors and factors relating to inner ear homeostasis. Various aquaporins (AQP), water channels, expressed in the inner ear and the vestibular organ, are involved in homeostasis. Mutations in AQP genes could result in disturbed inner ear homeostasis and endolymphatic hydrops, and therefore be involved in the pathogenesis of MD. Aim: To search for mutations in AQP1 to 4 in patients suffering from MD.

LIF promotes neurogenesis and maintains neural precursors in cell populations derived from spiral ganglion stem cells

BACKGROUND: Stem cells with the ability to form clonal floating colonies (spheres) were recently isolated from the neonatal murine spiral ganglion. To further examine the features of inner ear-derived neural stem cells and their derivatives, we investigated the effects of leukemia inhibitory factor (LIF), a neurokine that has been shown to promote self-renewal of other neural stem cells and to affect neural and glial cell differentiation. RESULTS: LIF-treatment led to a dose-dependent increase of the number of neurons and glial cells in cultures of sphere-derived cells. Based on the detection of developmental and progenitor cell markers that are maintained in LIF-treated cultures and the increase of cycling nestin-positive progenitors, we propose that LIF maintains a pool of neural progenitor cells. We further provide evidence that LIF increases the number of nestin-positive progenitor cells directly in a cell cycle-independent fashion, which we interpret as an acceleration of neurogenesis in sphere-derived progenitors. This effect is further enhanced by an anti-apoptotic action of LIF. Finally, LIF and the neurotrophins BDNF and NT3 additively promote survival of stem cell-derived neurons. CONCLUSION: Our results implicate LIF as a powerful tool to control neural differentiation and maintenance of stem cell-derived murine spiral ganglion neuron precursors. This finding could be relevant in cell replacement studies with animal models featuring spiral ganglion neuron degeneration. The additive effect of the combination of LIF and BDNF/NT3 on stem cell-derived neuronal survival is similar to their effect on primary spiral ganglion neurons, which puts forward spiral ganglion-derived neurospheres as an in vitro model system to study aspects of auditory neuron development.

Robust postmortem survival of murine vestibular and cochlear stem cells

Potential treatment strategies of neurodegenerative and other diseases with stem cells derived from nonembryonic tissues are much less subjected to ethical criticism than embryonic stem cell-based approaches. Here we report the isolation of inner ear stem cells, which may be useful in cell replacement therapies for hearing loss, after protracted postmortem intervals. We found that neonatal murine inner ear tissues, including vestibular and cochlear sensory epithelia, display remarkably robust cellular survival, even 10 days postmortem. Similarly, isolation of sphere-forming stem cells was possible up to 10 days postmortem. We detected no difference in the proliferation and differentiation potential between stem cells isolated directly after death and up to 5 days postmortem. At longer postmortem intervals, we observed that the potency of sphere-derived cells to spontaneously differentiate into mature cell types diminishes prior to the cells losing their potential for self-renewal. Three-week-old mice also displayed sphere-forming stem cells in all inner ear tissues investigated up to 5 days postmortem. In summary, our results demonstrate that postmortem murine inner ear tissue is suited for isolation of stem cells.

A Novel Implantable Hearing System with Direct Acoustic Cochlear Stimulation

A new implantable hearing system, the direct acoustic cochlear stimulator (DACS) is presented. This system is based on the principle of a power-driven stapes prosthesis and intended for the treatment of severe mixed hearing loss due to advanced otosclerosis. It consists of an implantable electromagnetic transducer, which transfers acoustic energy directly to the inner ear, and an audio processor worn externally behind the implanted ear. The device is implanted using a specially developed retromeatal microsurgical approach. After removal of the stapes, a conventional stapes prosthesis is attached to the transducer and placed in the oval window to allow direct acoustical coupling to the perilymph of the inner ear. In order to restore the natural sound transmission of the ossicular chain, a second stapes prosthesis is placed in parallel to the first one into the oval window and attached to the patient's own incus, as in a conventional stapedectomy. Four patients were implanted with an investigational DACS device. The hearing threshold of the implanted ears before implantation ranged from 78 to 101 dB (air conduction, pure tone average, 0.5-4 kHz) with air-bone gaps of 33-44 dB in the same frequency range. Postoperatively, substantial improvements in sound field thresholds, speech intelligibility as well as in the subjective assessment of everyday situations were found in all patients. Two years after the implantations, monosyllabic word recognition scores in quiet at 75 dB improved by 45-100 percent points when using the DACS. Furthermore, hearing thresholds were already improved by the second stapes prosthesis alone by 14-28 dB (pure tone average 0.5-4 kHz, DACS switched off). No device-related serious medical complications occurred and all patients have continued to use their device on a daily basis for over 2 years. Copyright (c) 2008 S. Karger AG, Basel.

Ultrastructural changes in otoconia of osteoporotic rats

The etiology of benign paroxysmal positional vertigo (BPPV) remains obscure in many cases and women are affected more often than men. A recent prospective study, performed in women >50 years of age suffering from recurrent BPPV, showed associated osteopenia or osteoporosis in a large percentage of these patients. These results suggested the possible relationship between recurrent BPPV and a decreased fixation of calcium in bone in women >50 years. To test this hypothesis, an experimental study was performed in adult female rats. Utricular otoconia of female rats in which osteopenia/osteoporosis was induced by bilateral ovariectomy (OVX) were compared to those of sham-operated adult females rats (SHAM), as control group. FIRST STUDY: The morphology of theutricles of OVX and SHAM rats was analyzed with scanning electron microscopy. In osteopenic/osteoporotic rats, the density of otoconia (i.e. the number of otoconia per unit area) was decreased (p = 0.036)and their size was increased (p = 0.036) compared to the control group. SECOND STUDY: To test the role of calcium turnover in such morphological changes, utricular otoconia of 2 other groups of OVX and SHAM rats, previously injected with calcein subcutaneously, were examined by conventional and epifluorescence microscopy. In epifluorescence microscopy, labeling with calcein showed no significant fluorescence in either group. This finding was interpreted as a lack of external calcium turnover into otoconia of adult female rats. The ultrastructural modifications of otoconia in osteopenic/osteoporotic female adult rats as well as the role of estrogenic receptors in the inner ear are discussed. The possible pathophysiological mechanisms which support the relationship between recurrent BPPV in women and the disturbance of the calcium metabolism of osteopenia/osteoporosis are debated.

A two-microphone noise reduction system for cochlear implant users with nearby microphones. Part II: Performance Evaluation

Users of cochlear implants (auditory aids, which stimulate the auditory nerve electrically at the inner ear) often suffer from poor speech understanding in noise. We evaluate a small (intermicrophone distance 7 mm) and computationally inexpensive adaptive noise reduction system suitable for behind-the-ear cochlear implant speech processors. The system is evaluated in simulated and real, anechoic and reverberant environments. Results from simulations show improvements of 3.4 to 9.3 dB in signal to noise ratio for rooms with realistic reverberation and more than 18 dB under anechoic conditions. Speech understanding in noise is measured in 6 adult cochlear implant users in a reverberant room, showing average improvements of 7.9–9.6 dB, when compared to a single omnidirectional microphone or 1.3–5.6 dB, when compared to a simple directional two-microphone device. Subjective evaluation in a cafeteria at lunchtime shows a preference of the cochlear implant users for the evaluated device in terms of speech understanding and sound quality.