Histopathology of the Clarion cochlear implant electrode positioner in a human subject.

A Silastic electrode positioner was introduced by the Advanced Bionics Corporation in 1999 and it was designed to achieve a perimodiolar position of the stimulating electrode. The positioner was voluntarily recalled in the United States in July 2002 due to an apparent higher risk of bacterial meningitis in patients in whom the electrode positioner had been placed. A detailed histopathologic study of the positioner in the human has not previously been published. The histopathologic findings in a 74-year-old woman who underwent bilateral cochlear implantation using the positioner are presented. Findings include a large track caused by the combined electrode and its positioner with considerable disruption of the basilar membrane and osseous spiral lamina. Although there was a fibrous sheath around the electrode and positioner at the cochleostomy in both ears, this fibrous sheath did not extend deeply into the cochlea except at the apical end of the electrode beyond the positioner. This resulted in a large fluid space around and between the positioner and electrode within the cochlea and presumably in fluid continuity with the cerebrospinal fluid space. Possible clinical implications are discussed.

Aortic dissection in a case of Turner's syndrome.

A case report is presented of an aortic dissection in a patient with Turner's syndrome that went undiagnosed. A thorough discussion of this fatal condition in this group of patients is included.

Histopathology of cochlear implants in humans.

The insertion of an intrascalar electrode array during cochlear implantation causes immediate damage to the inner ear and may result in delayed onset of additional damage that may interfere with neuronal stimulation. To date, there have been reports on fewer than 50 temporal bone specimens from patients who had undergone implantation during life. The majority of these were single-channel implants, whereas the majority of implants inserted today are multichannel systems. This report presents the histopathologic findings in temporal bones from 8 individuals who in life had undergone multichannel cochlear implantation, with particular attention to the type and location of trauma and to long-term changes within the cochlea. The effect of these changes on spiral ganglion cell counts and the correlation between speech comprehension and spiral ganglion cell counts were calculated. In 4 of the 8 cases, the opposite, unimplanted ear was available for comparison. In 3 of the 4 cases, there was no significant difference between the spiral ganglion cell counts on the implanted and unimplanted sides. In addition, in this series of 8 cases, there was an apparent negative correlation between residual spiral ganglion cell count and hearing performance during life as measured by single-syllable word recognition. This finding suggests that abnormalities in the central auditory pathways are at least as important as spiral ganglion cell loss in limiting the performance of implant users.

Innervation of supporting cells in the apical turns of the guinea pig cochlea is from type II afferent fibers.

The outer supporting cells in the apical turns of the guinea pig cochlea receive a dense innervation. Our previous study (Fechner et al. [1998] J. Comp. Neurol. 400:299-300) suggested that this innervation of the Deiters' and Hensen's supporting cells was not derived from efferent fibers of the olivocochlear bundle, but its origin has not been further specified. To test the hypothesis that the innervation was afferent in origin, we traced apical afferent fibers that were retrogradely labeled by extracellular injections of horseradish peroxidase. Labeled afferent fibers were of two types: type I fibers contacted inner hair cells, whereas type II fibers crossed the tunnel and contacted outer hair cells. Significantly, most of the type II fibers also formed branches to the outer supporting cells. Although a few olivocochlear efferent fibers formed such branches, counts indicated that the overwhelming majority of the branches were produced by type II afferent fibers. These branches were not produced by basal type II fibers. Apical type II fibers also differed from basal fibers by having shorter lengths, spiraling both apically and basally, and contacting all three rows of outer hair cells. These innervation differences suggest differences in the ways that information from outer hair cells is processed in the apex versus the base of the cochlea.

Prevalence and ultrastructural morphology of axosomatic synapses on spiral ganglion cells in humans of different ages.

Axosomatic synapses were found on human spiral ganglion cells (HSGCs). Ultrastructural characterization and calculation of the prevalence of these synapses were performed by electron microscopic semi-serial sections of both type I and type II HSGCs, in specimens from subjects of ages 1 day, 14 days, 21 years and 51 years. Synapses on type I HSGCs were extremely rare. In contrast, axosomatic synapses were present on approximately 50% of type II HSGCs of a young adult. This prevalence seemed to vary by age. Thus, no synapses were found in a 1-day old neonate, few in a 14-day old, and on approximately 15% of the type II SGCs from a 51-year old specimen. The origin of the nerve fibers synapsing on the type II HSGCs could not be determined. In view of the fact that some of the fibers projected from the intraganglionic spiral bundle, which is known to contain olivocochlear efferents, these fibers may represent an efferent pathway to the spiral ganglion. However, since there was morphological evidence of more than one type of nerve fiber synapsing on type II HSGCs, other neural origins must be considered. Although the physiological function of these synapses is unknown, they may mediate pre-synaptic neural modulation of the type II HSGCs at the level of the spiral ganglion.

Pattern of degeneration of the spiral ganglion cell and its processes in the C57BL/6J mouse.

Although degeneration of spiral ganglion cells has been described as a histopathologic correlate of hearing loss both in animals and humans, the pattern and sequence of this degeneration remain controversial. Degeneration of hair cells and of spiral ganglion cells and their dendritic processes was evaluated in the C57BL/6J mouse, in which there is a genetically determined progressive sensorineural loss starting in the high frequencies that is similar to the pattern commonly seen in the human. Auditory function was evaluated by brainstem evoked responses, and degeneration of hair cells, ganglion cells and their dendrites was evaluated histologically at 3, 8, 12 and 18 months of age. Progressive loss of auditory sensitivity was correlated with the loss of outer and inner hair cells and spiral ganglion cells and their dendritic processes. In addition, dendritic counts were consistently lower at a distal location in the osseous spiral lamina (i.e. near the organ of Corti) than at a proximal location (i.e. near the spiral ganglion), and the difference between the number of distal dendrites and the number of proximal dendrites tended to be greater with advancing age. These observations suggest an age-related progressive retrograde degeneration of spiral ganglion cells. Thus, in degenerating cochleas, some remaining spiral ganglion cells may have no distal dendritic processes near the organ of Corti. This may have implications for successful stimulation of the cochlear neuron in cochlear implantation.

Dense innervation of Deiters' and Hensen's cells persists after chronic deefferentation of guinea pig cochleas.

Innervation of Deiters' and Hensen's cells has been described in the organ of Corti of several mammalian species and has been suggested to arise from the olivocochlear (OC) efferent system (Wright and Preston [1976] Acta Otolaryngol. 82:41-47). In the present study, antineurofilament immunostaining was used to reveal these outer supporting cell fibers (OSCFs) in the normal guinea pig. In control ears, OSCFs were absent in the basal half of the cochlea but increased in number steadily toward the apex, peaking at values of over 1,200 fibers/mm. These values indicate a far more profuse innervation of supporting cells than has been described previously, suggesting that most OSCFs were not stained in previous immunohistochemical studies. Chronic cochlear deefferentation was used to test whether OSCFs are part of the OC system. The OC bundle was transected unilaterally, and the animals were allowed to survive for 4-8 weeks. Completeness of deefferentation was assessed by using acetylcholinesterase staining of the brainstem and measurement of the density of OC fascicles in the cochlea. By using these metrics, unilateral deefferentation was nearly complete in three animals. In successfully deefferented cases, the OSCF innervation density was not statistically different from control values. We conclude that the vast majority of OSCFs are not of OC origin. We speculate that they may be branches of type II afferent fibers to outer hair cells and that a smaller population of OSCFs with different morphology and immunoreactivity may arise from the OC system.

Morphologic evidence for innervation of Deiters' and Hensen's cells in the guinea pig.

The presence of nerve fibers and terminals among Deiters' and Hensen's cells of the organ of Corti of the adult guinea pig is demonstrated using immunostaining for synaptophysin and neurofilaments, acetylcholinesterase histochemistry, and transmission electron microscopy. These nerve terminals appeared to form chemical synapses with Deiters' and Hensen's cells. Nerve fibers and synapses were more common in the apical as compared to the basal cochlea. The terminals were often present on basal appendages of Hensen's cells, which were rich in mitochondria and often contained a Golgi apparatus and dense core vesicles. Electron microscopy and immunostaining for neurofilaments showed that most Hensen's cells in the apical cochlea received innervation. Few of the nerve fibers and terminals were positive for acetylcholinesterase, which suggests that they were not collaterals of cholinergic olivocochlear fibers. The density of these fibers, as shown by immunohistochemistry for neurofilaments, was far greater than previous reports of GABA-ergic fibers, which suggests that they were not GABA-ergic olivocochlear fibers. The role of such fibers and synapses with supporting cells of the outer hair cell area is unknown. Determination of the origins and functions of these fibers will provide new insights into cochlear structure and function.

Morphologic evidence for three cell types in the human spiral ganglion.

Although two types of spiral ganglion cells (large type I and smaller type II) have classically been described by anatomic studies in both animal and human spiral ganglion, there is physiologic and morphologic evidence for subtypes of the large type I ganglion cell. In addition, in the animal and human, a variety of morphologic differences based on cytoplasmic content, myelinization, immunostaining and morphometric analysis have suggested more than one variety of type I ganglion cell. Light and electron microscopic serial sections of the spiral ganglion in two human specimens in the basal, middle and upper middle turns were pooled for morphometric analysis of the cell area, nuclear area and axon diameter. Analysis of variance, bivariate scatter plots and multivariate cluster analysis provided evidence for 3 types of ganglion cells in the human spiral ganglion: large, intermediate and small, varying from each other significantly on the basis of cell area. It was suggested, based on the morphologic findings and prevalence of the cell types, that the large and intermediate cells were subtypes of the classic type I spiral ganglion cell, whereas the small ganglion cell was consistent with the classically described type II ganglion cell.

Patterns of degeneration in the human cochlear nerve.

The patterns of neural degeneration of the spiral ganglion were studied in 12 human pathologic specimens and 2 normal neonatal specimens. Morphometric analysis of spiral ganglion cells included the maximum cross-sectional areas of both large (type 1) and small (type II) spiral ganglion cells. The organ of Corti in segments corresponding to the spiral ganglion, was evaluated for the presence or absence of inner (IHC) and outer (OHC) hair cells and supporting cells. The relationship between degeneration of spiral ganglion cells and degeneration in the organ of Corti, the age, sex, duration of deafness, cochlear location and delay between death and fixation was evaluated statistically. Both primary and secondary degeneration of the spiral ganglion were more severe in the basal than apical half of the cochlea. Degeneration of the spiral ganglion was most severe when both IHCs and OHCs were absent in the organ of Corti. No survival advantage was identified for type II ganglion cells as has been previously reported. That is, there was no correlation between the degree of degeneration of the spiral ganglion and the prevalence of type II ganglion cells. In fact, there was more severe degeneration of type II cells when the corresponding organ of Corti was severely degenerated. These findings in the human were compared with animal models of degeneration of the spiral ganglion, and the implications for cochlear implantation were discussed.

Supranuclear efferent synapses on outer hair cells and Deiters' cells in the human organ of Corti.

Synaptophysin immunoreactivity and transmission electron microscopy have demonstrated vesiculated nerve endings synapsing on the supranuclear zone of outer hair cells and also of Deiters' cells in the human organ of Corti. These fibers seem similar to supranuclear fibers, apparently derived from the olivocochlear efferent system, which have been described in the animal. However, these endings were found throughout the cochlea in the human whereas in the animal such fibers were limited to the apical cochlea. Although such fibers have been demonstrated among supporting cells by immunohistochemical techniques, this is the first demonstration by transmission electron microscopy of morphology consistent with a chemical synapse between such fibers and Deiters' cells. Although the role of such fibers is unknown, neurophysiologic evidence suggests that they may modify the micromechanics of the outer hair cell. The function of neural innervation of Deiters' cells is speculative.

Otopathology in a case of multichannel cochlear implantation.

The histopathology of the temporal bones of a patient who died of unrelated causes 10 weeks following cochlear implantation using a Richards Ineraid device is presented. Deafness was caused by a prolonged course of intravenous gentamycin therapy 5 years prior to implantation. The electrode array of the cochlear implant was left in situ throughout histologic preparation and sectioning. Despite displacement and disruption of supporting structures of the inner ear, particularly in the 6-to-15-mm range as measured from the round window, there was no significant difference in the mean densities of spiral ganglion cells in the implanted and unimplanted sides. This case is presented as evidence that despite significant disruption of supporting elements of the inner ear, which is common during cochlear implantation, there appears to be little effect on the residual spiral ganglion cell count, at least in the short term.

Proteolytic cleavage at arginine residues within the hydrophilic disulphide loop of the Escherichia coli Shiga-like toxin I A subunit is not essential for cytotoxicity.

Escherichia coli Shiga-like toxin I is a type II ribosome-inactivating protein composed of an A subunit with RNA-specific N-glycosidase activity, non-covalently associated with a pentamer of B subunits possessing affinity for galabiose-containing glycolipids. The A subunit contains a single intrachain disulphide bond encompassing a hydrophilic sequence containing two trypsin-sensitive arginine residues. By analogy with other bacterial toxins it has been proposed that proteolytic nicking, deemed essential for a cytotoxic effect, occurs within this disulphide-bonded loop to generate the A1 and A2 fragments. Reduced A1 is then believed to translocate an internal membrane to inactivate protein synthesis in the cytosol. In this report, the disulphide-loop arginines of the SLT I A subunit were mutated to block the specific proteolysis presumed to occur. However, the mutant generated remained an effective toxin having similar catalytic activity to wild-type toxin and only a marginally reduced cytotoxicity towards cultured cells. We conclude that the disulphide-loop arginine residues are not the unique and essential processing sites previously assumed, but that processing may occur at alternative accessible sites to compensate for loss of target sites within the loop.

The localization of synaptophysin in the organ of Corti of the human as shown by immunoelectron microscopy.

Synaptophysin, or p38, a polypeptide of molecular weight 38 kD, is a calcium-binding membrane protein found in synaptic vesicles of neurons and smooth surfaced vesicles of neuroendocrine cells. Six human neonatal and infant temporal bones were fixed in paraformaldehyde and glutaraldehyde, decalcified in EDTA and were than immunoreacted for synaptophysin (ICN Biomedicals) using the avidin-biotin reaction (ABC kit, Vector Labs). The tissue was then prepared for light microscopic surface preparation, radial sections of 5 microns, and serial section electron microscopy. At a light microscopic level, the inner spiral bundle, tunnel spiral bundle, upper tunnel crossing fibers and the base of outer hair cells were stained. At the base of outer hair cells, the immunoreactivity was seen to decrease from the base to the apex and from the first to third outer hair cells. At an electron microscopic level, immunoreactivity at the base of outer hair cells was limited to vesiculated efferent fibers. The degree of immunoreactivity between adjacent efferent fibers varied significantly. Immunoreactive vesiculated endings were also found in the supranuclear region of outer hair cells.

Postnatal maturation of human spiral ganglion cells: light and electron microscopic observations.

The presence of two types of ganglion cells, based on cell size and other morphologic parameters, is well established in the adult mammalian and human spiral ganglion. On the other hand, there is little data concerning cell morphology in the neonatal spiral ganglion. The present study was undertaken to evaluate the differences in the morphometry and distribution of cell types in the spiral ganglion of the human neonate as compared to the adult. A total of five human temporal bones from two neonates and three infants were included in this study. Light microscopic analysis of all specimens was performed, and electron microscopic evaluation of a 14 day old neonatal spiral ganglion was accomplished. The segmental density of spiral ganglion cells was higher in the neonate than in the adult. The prevalence of type II spiral ganglion cells was higher in the neonate than has been reported in the adult, particularly in the middle and apical turns where type II cells constituted 24% and 26% of all ganglion cells, respectively. The prevalence of type II ganglion cells decreased with age, particularly in the middle and apical turns. In the neonate, the maximal cross sectional area of type I neurons increased from the base to the apex and seemed to increase with age especially in the basal turn. The present study strongly supports a clear differentiation of type I and type II ganglion cells in the human neonate and that the prevalence of type II cells is greater in the neonate than the adult. This finding is discussed with reference to postnatal development of the spiral ganglion.

Morphometric analysis of normal human spiral ganglion cells.

A morphometric analysis of the human spiral ganglion was performed at an ultrastructural level. Cells of the basal, middle, and upper middle turns were evaluated by serial section. The following parameters were evaluated: myelinization, area, diameter, circumference, and degree of roundness of the cell body and nucleus; diameter of the axon and dendrite in their initial segments; and process configuration. Analysis of variance suggested there were at least two types of cell bodies within the human spiral ganglion, best delineated by the dimensions of the cell body and nucleus and the ratio of diameter of the initial segments of axonic and dendritic processes. Myelinization and degree of roundness were relatively unimportant differential parameters. Cluster (multivariate) analysis of the six most important differential parameters suggested the possibility of five subgroups among the groups of large and small cells. For the middle and upper middle turns, cluster analysis suggested that there may be as many as three groups of cells based on morphometric analysis. These results are discussed in relation to other morphologic and physiologic data in the mammalian spiral ganglion.

Morphology of synapses at the base of hair cells in the organ of Corti of the chimpanzee.

The synaptic morphology of inner and outer hair cells of the organ of Corti of the chimpanzee was evaluated by serial section electron microscopy. The morphology of nerve terminals and synapses at both sites was very similar to that of human and other mammalian species. Two types of nerve terminals, nonvesiculated and vesiculated, with distinct synaptic morphology were found. In addition, between some nonvesiculated endings and outer hair cells, a reciprocal synaptic relationship was seen. In such terminals there was morphologic evidence for transmission from hair cell to neuron and from neuron to hair cell between a single neuron and an outer hair cell.

Comparison of an inductively coupled plasma-atomic emission spectrometry method for the determination of calcium, magnesium, sodium, potassium, copper and zinc with atomic absorption spectroscopy and flame photometry methods.

Serum calcium, magnesium, copper and zinc concentrations obtained from the analysis of 77 serum samples by inductively coupled plasma-atomic emission spectrometry (ICP-AES) are compared with the results obtained using atomic absorption spectroscopy (AAS). Similarly, serum sodium and potassium levels from the analysis of the same samples by ICP-AES are compared with the levels obtained by flame photometry. For each metal, we compare the results from both methods with a linear regression program that assumes error in both variables. The regression analysis shows that the ICP-AES method gives slightly higher calcium, copper, and zinc results and lower magnesium results than the AAS methods, and lower sodium and potassium results than the flame photometry method. Except for sodium, the correlation (r) between the results is very high (greater than or equal to 0.958), indicating that the ICP-AES results could be corrected to be equivalent to the atomic absorption or flame photometry results. The ICP-AES has the advantage of requiring less preparation and analysis time, and additional elements could be determined simultaneously in the same sample.