5,772 research outputs found
A mechanism for sensing noise damage in the inner ear
No full textOur sense of hearing requires functional sensory hair cells. Throughout life those hair cells are subjected to various traumas, the most common being loud sound. The primary effect of acoustic trauma is manifested as damage to the delicate mechanosensory apparatus of the hair cell stereocilia [1]. This may eventually lead to hair cell death [2] and irreversible deafness [3]. Little is known about the way in which noxious sound stimuli affect individual cellular components of the auditory sensory epithelium. However, studies in different types of cell cultures have shown that damage and mechanical stimulation can activate changes in intracellular free calcium concentration ([Ca(2+)](i)) and elicit intercellular Ca(2+) waves [4]. Thus an attractive hypothesis is that changes in [Ca(2+)](i), propagating as a wave through support cells in the organ of Corti, may constitute a fundamental mechanism to signal the occurrence of hair cell damage. The mechanism we describe here exhibits nanomolar sensitivity to extracellular ATP, involves regenerative propagation of intercellular calcium waves due to ATP originating from hair cells, and depends on functional IP(3)-sensitive intracellular stores in support cells
Permeability and Gating Properties of Human Connexins 26 and 30 expressed in HeLa cells.
No full textHuman connexins 26 and 30 were expressed either through the bicistronic pIRES-EGFP expression vector or as EYFP-tagged chimeras. When transiently transfected in communication-incompetent HeLa cells, hCx26–pIRES transfectants were permeable to dyes up to 622 Da, but were significantly less permeable to 759 Da molecules. Under the same conditions, permeability of hCx26–EYFP fusion products was comparable to that of hCx26–pIRES, but with significant increase in diffusion at 759 Da, possibly as a consequence of having selected large fluorescent junctional plaques. Dye transfer was limited to 457 Da in hCx30–EYFP transfectants. When reconstructed from confocal serial sections, fluorescent plaques formed by hCx26–EYFP and hCx30–EYFP appeared irregular, often with long protrusions or deep invagination. Similar plaques were observed following immunostaining both in cells transfected with hCx26–pIRES and in HeLa cells stably transfected with mouse Cx26. Tissue conductance (Tgj) displayed significantly smaller values (28.8 ± 1.8 nS) for stably transfected mCx26 than transiently transfected hCx26 (43.5 ± 3.3 nS). These differences reflected in distinct functional dependence of normalized junctional conductance (Gj) on transjunctional voltage (Vj). The half-activation voltage for Gj was close to ±95 and ±58 mV in mCx26 and hCx26, respectively. The corresponding parameters for hCx30 transfectants were Tgj=45.2±3.5 nS and V0=±34 mV. These results highlight unexpected differences between mCx26 and hCx26 in this expression system, reinforce the concept that channel permeability may be related to Cx level expression, and indicate that fusion of hCx30 to GFP colour mutants produces channels that are suitable for permeability and gating studies
Multiphoton Fluorescence Microscopy with GRIN Objective Aberration Correction by Low Order Adaptive Optics
No full textGraded Index (GRIN) rod microlenses are increasingly employed in the assembly of optical probes for microendoscopy applications. Confocal, two-photon and optical coherence tomography (OCT) based on GRIN optical probes permit in-vivo imaging with penetration depths into tissue up to the centimeter range. However, insertion of the probe can be complicated by the need of several alignment and focusing mechanisms along the optical path. Furthermore, resolution values are generally not limited by diffraction, but rather by optical aberrations within the endoscope probe and feeding optics. Here we describe a multiphoton confocal fluorescence imaging system equipped with a compact objective that incorporates a GRIN probe and requires no adjustment mechanisms. We minimized the effects of aberrations with optical compensation provided by a low-order electrostatic membrane mirror (EMM) inserted in the optical path of the confocal architecture, resulting in greatly enhanced image quality
Dissecting key components of the Ca2+ homeostasis game by multi-functional fluorescence imaging
No full textDifferent sub-cellular compartments and organelles, such as cytosol, endoplasmic reticulum and mitochondria, are known to be differentially involved in Ca2+ homeostasis. It is thus of primary concern to develop imaging paradigms that permit to make out these diverse components. To this end, we have constructed a complete system that performs multi-functional imaging under software control. The main hardware components of this system are a piezoelectric actuator, used to set objective lens position, a fast-switching monochromator, used to select excitation wavelength, a beam splitter, used to separate emission wavelengths, and a I/O interface to control the hardware. For these demonstrative experiments, cultured HeLa cells were transfected with a Ca2+ sensitive fluorescent biosensor (cameleon) targeted to the mitochondria (mtCam), and also loaded with cytosolic Fura2. The main system clock was provided by the frame-valid signal (FVAL) of a cooled CCD camera that captured wide-field fluorescence images of the two probes. Excitation wavelength and objective lens position were rapidly set during silent periods between successive exposures, with a minimum inter-frame interval of 2 ms. Triplets of images were acquired at 340, 380 and 430 nm excitation wavelengths at each one of three adjacent focal planes, separated by 250 nm. Optical sectioning was enhanced off-line by applying a nearest-neighbor deconvolution algorithm based on a directly estimated point-spread function (PSF). To measure the PSF, image stacks of sub-resolution fluorescent beads, incorporated in the cell cytoplasm by electroporation, were acquired under identical imaging conditions. The different dynamics of cytosolic and mitochondrial Ca2+ signals evoked by histamine could be distinguished clearly, with sub-micron resolution. Other FRET-based probes capable of sensing different chemical modifications of the cellular environment can be integrated in this approach, which is intrinsically suitable for the analysis of the interactions and cross-talks between different signaling pathways (e.g. Ca2+ and cAMP)
Development and subunit composition of synaptic NMDA receptors in the amygdala: NR2B Synapses in the adult central amygdala
No full textNMDA receptors are well known to play an important role in synaptic development and plasticity. Functional NMDA receptors are heteromultimers thought to contain two NR1 subunits and two or three NR2 subunits. In central neurons, NMDA receptors at immature glutamatergic synapses contain NR2B subunits and are largely replaced by NR2A subunits with development. At mature synapses, NMDA receptors are thought to be multimers that contain either NR1/NR2A or NR1/NR2A/NR2B subunits, whereas receptors that contain only NR1/NR2B subunits are extrasynaptic. Here, we have studied the properties of NMDA receptors at glutamatergic synapses in the lateral and central amygdala. We find that NMDA receptor-mediated synaptic currents in the central amygdala in both immature and mature synapses have slow kinetics and are substantially blocked by the NR2B-selective antagonists (1S, 2S)-1-(4-hydroxyphenyl)-2-(4-hydroxy-4-phenylpiperidino)-1-propano and ifenprodil, indicating that there is no developmental change in subunit composition. In contrast, at synapses on pyramidal neurons in the lateral amygdala, whereas NMDA EPSCs at immature synapses are slow and blocked by NR2B-selective antagonists, at mature synapses their kinetics are faster and markedly less sensitive to NR2B-selective antagonists, consistent with a change from NR2B to NR2A subunits. Using real-time PCR and Western blotting, we show that in adults the ratio of levels of NR2B to NR2A subunits is greater in the central amygdala than in the lateral amygdala. These results show that the subunit composition synaptic NMDA receptors in the lateral and central amygdala undergo distinct developmental changes
Author Co-Citation Analysis (ACA): a powerful tool for representing implicit knowledge of scholar knowledge workers
No full textIn the last decade, knowledge has emerged as one of the most important and valuable organizational assets. Gradually this importance caused to emergence of new discipline entitled ―knowledge management‖. However one of the major challenges of knowledge management is conversion implicit or tacit knowledge to explicit knowledge. Thus Making knowledge visible so that it can be better accessed, discussed, valued or generally managed is a long-standing objective in knowledge management. Accordingly in this paper author co- citation analysis (ACA) will be proposed as an efficient technique of knowledge visualization in academia (Scholar knowledge workers)
Portfolio of recorded performances and exegesis: Messiaen’s musical language for the jazz pianist - an exploration through performance.
No full textMoving beyond Gunther Schuller’s Third Stream amalgamation of classical and jazz, this study explores whether the musical language of Olivier Messiaen can make a valid contribution to jazz piano performance. Initially, my project sought to answer such questions as: What elements of the musical language of Messiaen already exist in the jazz vocabulary? Am I able to extend this further? What are the timbral structures and pianistic effects within Messiaen’s musical language? What will be the most effective application of Messiaen’s musical language to jazz piano performance? Endeavouring to answer the final question led me to consider such aspects as whether the project should be limited to quoting Messiaen motifs, arranging Messiaen melodies, replacing jazz harmonic structures on standards with examples from Messiaen’s musical language or whether it would be better to approach the research conceptually. The work of Hubert Nuss provided encouraging reassurance that this was not an impossible task. In order to articulate this conception, the initial challenge was to decide how the classical and jazz worlds might meet in a ‘Messiaen’ technique. The approach adopted was similar to that used for undergraduate jazz study, namely, immersion in the piano scores and recordings of Messiaen’s music as well as by live performances. This was followed by the development and assessment of a contrived approach when specific techniques, such as tonal colourings or harmonic structures, were developed through prepared exercises and consciously included in my performance. It was then compared with an intuitive approach when no such precise parameters were established. This submission consists of CD recordings of two public recitals and an exegesis. It documents the development of this Messiaen technique and discusses its application in my performances. It also demonstrates the ways that Messiaen’s musical language can be used within jazz piano performance to provide a colour that distinguishes jazz piano performance in a competitive field.Thesis (M.Mus.) -- University of Adelaide, Elder Conservatorium of Music, 201
Spinal anaesthesia for brachytherapy for carcinoma of the cervix a comparison of two dose regimes of hypebaric bupivacaine
No full textIncludes abstract.Includes bibliographical references.The main purpose of the study was to help establish the best dose regimen of hyperbaric bupivacaine, when combined with intrathecal fentanyl, for spinal anaesthesia for brachytherapy for carcinoma of the cervix. This procedure is performed as a day case at Groote Schuur Hospital
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