Michael L. Wiederhold

Experimental Long-term Tubal Occlusion in Cats: A Quantitative Histopathological Study

Unilateral tubal occlusion was performed in 10 cats, which were sacrificed 26-44 months later. During the observation period repeated bilateral tympanometry and auditory nerve response were recorded. Paracentesis with evacuation of effusion was performed up to eight times and the effusion was examined by viscometry. After sacrifice, quantitative histopathology of the bulla with determination of goblet cell density was carried out. In three ears having glue at the last paracentesis, the histopathologic changes were more pronounced than in the four ears having thin effusion. Three ears were dry at sacrifice with normal tympanometry, the histopathological changes evidently demonstrate the the normalization of the middle ear mucosa. The experiments demonstrate the great importance of tubal patency or function in prognosis of the disease. In ears with anatomically occluded tubes, no recovery occurred, whereas in ears with normalized tubal function, the middle ear mucosa and goblet cell density also normalized. Pathogenic and etiological aspects of secretory otitis are discussed.

High-frequency electrostimulation hearing after mastoidectomy.

This study was undertaken to measure the degree of high-frequency sensorineural hearing loss following mastoid surgery. Twenty-five patients undergoing mastoidectomy procedures were tested preoperatively, less than 2 days postoperatively, and at 30 days postoperatively using the Tonndorf Audimax 500 high-frequency audiometer. Electrostimulation thresholds in 1-kHz intervals, from 1 to 20 kHz, were measured, and the highest detectable frequency was determined to within 0.1 kHz. Surgical drilling time was recorded. Average drilling time was 51 minutes. A significant temporary threshold shift was observed, measurable at multiple frequencies, less than 48 hours after mastoidectomy. There was no clinically significant change in electrostimulation thresholds (measured in 1-kHz increments, from 1 to 16 kHz) preoperatively to 30 days postoperatively. A statistically significant average loss of 0.89 kHz in the highest frequency producing a measurable response was noted (p < 0.05). Determinations of the highest measurable frequency may be the most sensitive measure of surgically-induced, high-frequency sensorineural hearing changes.

Acoustic Overstimulation Reduces 2f1-f2 Cochlear Emissions at All Levels in the Cat

The cubic distortion product, 2f1 — f2 (DP), was recorded in the cat ear canal as the levels of two primary tones (f1=4.0, f2=5.2 kHz here) were varied from approximately 40 to 90dB SPL. The DP level grows as the cube of the primary level at low and high levels, and usually exhibits a dip near L1 = 65dB SPL, DP growth functions and N1 amplitude-level functions were compared before and after exposure to either 4 kHz pure tones or to 500 Hz octave bands of noise. Pure-tone exposure produced a dose-dependent shift in the N1 growth whereas the low-frequency noise exposure produced widely varying shifts in N1 functions. In either case, however, changes in DP were correlated with changes in the neural responses. The level of the DP was compared before and after exposure at 2 levels below and one above the dip at L1 = 65dB SPL. For all of these measures, the decrease in DP level was significantly correlated with the shift in the N1 growth function. At both low and high primary levels, decrease in DP grows approximately as the square of the N1 shift for shifts greater than 15dB.

A mechanism of adaptation to hypergravity in the statocyst of Aplysia californica

The gravity-sensing organ of Aplysia californica consists of bilaterally paired statocysts containing statoconia, which are granules composed of calcium carbonate crystals in an organic matrix. In early embryonic development, Aplysia contain a single granule called a statolith, and as the animal matures, statoconia production takes place. The objective of this study was to determine the effect of hypergravity on statoconia production and homeostasis and explore a possible physiologic mechanism for regulating this process. Embryonic Aplysia were exposed to normogravity or 3 x g or 5.7 x g and each day samples were analyzed for changes in statocyst, statolith, and body dimensions until they hatched. In addition, early metamorphosed Aplysia (developmental stages 7-10) were exposed to hypergravity (2 x g) for 3 weeks, and statoconia number and statocyst and statoconia volumes were determined. We also determined the effects of hypergravity on statoconia production and homeostasis in statocysts isolated from developmental stage 10 Aplysia. Since prior studies demonstrated that urease was important in the regulation of statocyst pH and statoconia formation, we also evaluated the effect of hypergravity on urease activity. The results show that hypergravity decreased statolith and body diameter in embryonic Aplysia in a magnitude-dependent fashion. In early metamorphosed Aplysia, hypergravity decreased statoconia number and volume. Similarly, there was an inhibition of statoconia production and a decrease in statoconia volume in isolated statocysts exposed to hypergravity in culture. Urease activity in statocysts decreased after exposure to hypergravity and was correlated with the decrease in statoconia production observed. In short, there was a decrease in statoconia production with exposure to hypergravity both in vivo and in vitro and a decrease in urease activity. It is concluded that exposure to hypergravity downregulates urease activity, resulting in a significant decrease in the formation of statoconia.

Spinner cephalopods: defects of statocyst suprastructures in an invertebrate analogue of the vestibular apparatus.

SummaryIndividuals of seven species of coleoid cephalopods (three species of octopus, three of squid, and one of cuttlefish), that were cultured and reared in laboratory aquarium systems, had a behavioral defect at hatching which was characterized by an inability to control orientation while swimming. These defective animals were designated as “spinners.”An examination of statocysts from individuals of five of the affected species revealed abnormalities of the neuroepithelial suprastructures: absence or malformation of the statolith of the gravity receptor system and absence of the cupulae of the angular acceleration receptor systems. The sensory epithelia did not differ from those of normal animals, nor did the synaptic structures and relationships, when examined both with scanning and transmission electron microscopy. The abnormalities were compared with congenital defects of the neuropeithelial suprastructures of the vestibular apparatus (especially in mammals). The defects observed in statocysts of spinner animals are thought to be the result of environmental causes, such as the temperature or chemistry of the seawater in the transportation vessels or rearing systems, rather than genetic causes.

Development of the otolith organs and semicircular canals in the Japanese red-bellied newt, Cynops pyrrhogaster

The sequence in which the otoliths and semicircular canals and their associated sensory epithelia appear and develop in the newt are described. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos from stages 31 through 58 demonstrate the first appearance, relative position and growth of the otoliths. A single otolith is first seen in stage 33 embryos (approximately 9 days old); this splits into separate utricular and saccular otoliths at stage 40 (13 days). The lateral semicircular canal is the first to appear, at stage 41 (14 days). The anterior and posterior canals appear approximately one week later and the vestibular apparatus is essentially fully formed at stage 58 (approximately 5 weeks). The data reported here will serve as ground-based controls for fertilized newt eggs flown on the International Microgravity Laboratory-2 Space Shuttle flight, to investigate the influence of microgravity on the development of the gravity-sensing organs.

Visualization of newt aragonitic otoconial matrices using transmission electron microscopy

Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. These protein matrices are thought to originate from the supporting or hair cells in the macula during development. Previous studies of mammalian calcitic, barrel-shaped otoconia revealed an organized protein matrix consisting of a thin peripheral layer, a well-defined organic core and a flocculent matrix inbetween. No studies have reported the microscopic organization of the aragonitic otoconial matrix, despite its protein characterization. Pote et al. (1993b) used densitometric methods and inferred that prismatic (aragonitic) otoconia have a peripheral protein distribution, compared to that described for the barrel-shaped, calcitic otoconia of birds, mammals, and the amphibian utricle. By using tannic acid as a negative stain, we observed three kinds of organic matrices in preparations of fixed, decalcified saccular otoconia from the adult newt: (1) fusiform shapes with a homogenous electron-dense matrix; (2) singular and multiple strands of matrix; and (3) more significantly, prismatic shapes outlined by a peripheral organic matrix. These prismatic shapes remain following removal of the gelatinous matrix, revealing an internal array of organic matter. We conclude that prismatic otoconia have a largely peripheral otoconial matrix, as inferred by densitometry.

The morphogenic features of otoconia during larval development of Cynops pyrrhogaster, the Japanese red-bellied newt

Otoconia are calcified protein matrices within the gravity-sensing organs of the vertebrate vestibular system. Mammalian otoconia are barrel-shaped with triplanar facets at each end. Reptilian otoconia are commonly prismatic or fusiform in shape. Amphibians have all three otoconial morphologies, barrel-shaped otoconia within the utricle, with prismatic and fusiform otoconia in the saccule. Scanning electron microscopy revealed a sequential appearance of all three otoconial morphologies during larval development of the newt, Cynops pyrrhogaster. The first otoconia appear within a single, developing otolith, and some resemble adult barrel-shaped otoconia. As the larvae hatch, around stages 39-42, the single otolith divides into two anatomically separate regions, the utricle and saccule, and both contain otoconia similar to those seen in the single otolith. Throughout development, these otoconia may have variable morphologies, with serrated surfaces, or circumferential striations with either separated facets or adjacent facets in the triplanar end-regions. Small fusiform otoconia occur later, at stage 51, and only in the saccule. Prismatic otoconia appear later still, at stage 55, and again only in the saccule. Thus, although prismatic otoconia are the most numerous in adult newts, it is the last vestibular otoconial morphology to be expressed.

Contractile properties of laryngeal muscles in young and old baboons

Aging is associated with changes in voice and/or laryngeal protective reflexes in humans. To assess the role of the laryngeal muscles in this change, we have examined in vitro thyroarytenoid (vocalis) muscles excised from the larynxes of young and old baboons. Contractile properties, fatigue characteristics, and resting oxygen consumption of these muscles were measured. The results indicate that the thyroarytenoid muscle of the baboon is a fast muscle that is resistant to fatigue. The thyroarytenoid muscles of older baboons contracted more slowly and recovered less rapidly from fatigue induced by prolonged contractions but developed more maximum active tension than thyroarytenoid muscles of young adult baboons. These small differences in the characteristics of thyroarytenoid muscle of older baboons are probably not sufficient to explain observed changes in laryngeal function in the elderly. The results suggest that changes in neural pathways and/or morphology of the larynx may play a larger role in the altered laryngeal function with age.

Effects of hypergravity on statocyst development in embryonic Aplysia californica

Aplysia californica is a marine gastropod mollusc with bilaterally paired statocysts as gravity-receptor organs. Data from three experiments in which embryonic Aplysia californica were exposed to 2 x g are discussed. The experimental groups were exposed to excess gravity until hatching (9-12 day), whereas control groups were maintained at normal gravity. Body diameter was measured before exposure to 2 x g. Statocyst, statolith and body diameter were each determined for samples of 20 embryos from each group on successive days. Exposure to excess gravity led to an increase in body size. Statocyst size was not affected by exposure to 2 x g. Statolith size decreased with treatment as indicated by smaller statolith-to-body ratios observed in the 2 x g group in all three experiments. Mean statolith diameter was significantly smaller for the 2 x g group in Experiment 1 but not in Experiments 2 and 3. Defective statocysts, characterized by very small or no statoliths, were found in the 2 x g group in Experiments 1 and 2.