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Biomedical subjects

M Anniko

Publications and source records attributed to M Anniko.

At least 73 records · Page 4Linked to original sources

Electrophysiological effects of the clinically used local anesthetics lidocaine, lidocaine-prilocaine and phenol on the rat's inner ear.

Local anesthetics, even if applied to the outer ear canal, may still enter the middle ear, running the risk of penetrating the round window. To elucidate the effect of certain topical anesthetics on the inner ear, the round window niche in the laboratory rat's middle ear was exposed separately to lidocaine, lidocaine-prilocaine and phenol. Auditory brain-stem responses (ABR) were recorded at 2, 4, 6, 8, 12, 16, 20, and 31.5 kHz before the application, and 24 h, 3 weeks, 2 months and 6 months after exposure. After terminating the 6-month ABR measurements, the animal were sacrificed and the temporal bones fixed and decalcified for light microscopic analysis. All three drugs affected the ABR thresholds and the cochlear morphology with a pattern characteristic for each drug. At 24 h, all three substances caused severe impairment of ABR thresholds, followed by a period of restitution lasting up to 2 months. Even 6 months after exposure, the ABR thresholds at and above 12 kHz were impaired, as compared with the pretreatment level, for all substances tested. In the lower frequencies the original ABR threshold was reached in the order: (1) lidocaine, (2) lidocaine-prilocaine, (3) phenol. The cochlear structures were unaffected by lidocaine, whereas lidocaine-prilocaine and phenol caused morphological damage which was most pronounced after exposure to phenol. The heterogeneity of the changes in the ABR thresholds suggests differences in the mechanism of action of each type of local anesthetic investigated. The effects following lidocaine were transient. However, clinicians must be aware of the ototoxic potential of both lidocaine and phenol.

Anesthetics, Local↗

Morphologically specific vestibular hair cell degeneration in the jerker mouse mutant.

The vestibular organs in homozygote (je/je) and heterozygote (je/+) 1- to 12-month-old jerker mouse mutants were analyzed morphologically using light microscopy and transmission and scanning electron microscopy. The pathological changes were initially confined to the surface structure of predominantly type I hair cells. The latter cells showed disintegration of the cuticular plate, disarray of stereocilia and finally fusion of sensory hairs. The hair cells were then expelled into the endolymphatic space. A notable finding was that most expelled hair cells still showed a rather normal ultrastructure of their cytoplasm and nuclei. The morphological type of pathology was the same in je/je and je/+ mutants. However, in je/je animals the degeneration occurred within 1-3 months after birth, whereas in je/+ mutants a slowly progressive degeneration lasted for up to 1 year of age or even more. The degeneration was more extensive in the two maculae than in the three cristae ampullares.

Aging↗

[The cytokeratin skeleton of the human organ of Corti and its functional significance].

In the adult human organ of Corti cytokeratin (CK) is expressed by all supporting cells enclosing it like a shell. The pattern of immunoreactivity clearly demonstrates a quantitative gradient in the expression of CK, with more CK at the apex than at the base of the cochlea. Predominantly in the apical cochlear turns, the CK-shell separates the compartment of the inner hair cells from that of the outer hair cells. Ultrastructurally, the supporting cells contain a loose fibrillary network which apically is oriented toward the desmosome chain and which can be clearly distinguished from the well-known tubular filaments (microtubuli). Some supporting cells show centrioles. Both the expression of CK and the presence of centrioles indicate a possible potential for cell regeneration. Ultrastructurally, outer hair cells and Deiters' cells show features of a specialized contact zone which might be of functional significance regarding the contractile abilities of the outer hair cells. In the human organ of Corti, vimentin is expressed only by the inner and outer pillar cells. These cells thus express CK together with vimentin. The distinct shell configuration of the CK network in the organ of Corti gives it a tonotopically related difference in rigidity which not only must be of importance for cochlear perception of sound but also could explain the reduced vulnerability of the ear to low frequencies.

Cytoskeleton↗

[Functional morphology of the outer hair cells of the human: new aspects].

Serial sections through the outer hair cells of the human organ of Corti were investigated using high-resolution electron microscopy. Delicate monostratified tubular structures of differing lengths are located on the cytoplasmic side of the hair-cell membrane. They are connected to the cell membrane via short fibers (pillars) arranged in pairs. The tubular structures, which are also termed subsurface cisterns (SSC), constitute an extensive network. This network lines the entire inner surface of the outer hair-cell membrane, and in contrast to the mammals investigated so far, also lines the base of the cell. The postsynaptic cisterns of the efferent synapses are integrated into this system. At the surface of the inner lamina of the cisterns, pores 8.5 nm in diameter are to be found. These are surrounded by protein complexes with a stellate arrangement. Similar protein complexes are located on the inner lamina of the postsynaptic cisterns. Pores 8.5 nm in diameter are also present in the outer lamina of the SSC. These constitute the base for the pillar filaments. At high magnification, the pillars are seen to have a lumen about 6 nm in diameter. Pores about 8 nm in diameter are again to be found where the pillars are anchored in the outer cell membrane. The pillars, consisting of actin, are probably ionic channels, and the pores of the inner lamina of the SSC surrounded by protein complexes are thought to be acetylcholine receptors. Since the postsynaptic cisterns of the efferent innervation of the outer hair cells are part of the overall system of the SSC, and acetylcholine is regarded as the neurotransmitter of efferent innervation, it would seem possible to regulate the subsurface cisterns, as a wall-stabilizing, contractile system, via the efferent innervation. On the basis of serial sections, a three-dimensional, true-to-scale model of the contractile wall system of the human outer hair cell was reconstructed.

Adult↗

Sealing of the round window reversibly affects auditory brainstem response latencies.

The round window (RW) niche was sealed in 12 adult female albino Sprague-Dawley rats. Electrophysiological auditory function was measured with a computerized auditory brainstem response (ABR) recording technique, using 1- and 6 kHz stimuli. ABR measurements were performed prior to closure of the niche, immediately after the sealing, and 4 h, 24 h and 7 days later. After sealing, the latencies elicited by a 6 kHz stimulus increased, but returned to normal between 24 h and 7 days after closure. Stimulation with 1 kHz did not influence the latencies in such a clear-cut manner as when using 6 kHz. It is conceivable that sealing of the RW deprives a minor fraction of oxygen supplied to the basal part of the cochlea through the RW membrane, a procedure which by us is monitored as transient electrophysiological changes.

Animals↗

Long-term survival of patients with paranasal sinus carcinoma.

The survival rate for all patients with a squamous cell carcinoma of the paranasal sinuses (n = 77) diagnosed in 1960-1985 in a population of approximately 1 million inhabitants was analysed without any selection of cases. This population is geographically restricted to the most northern part of Sweden. The 5-year survival rates of patients with T3 and T4 carcinomas were 42 and 14%, respectively (corrected survival), and the 10-year survival rates were 40 and 14%, respectively. In patients with tumours which initially were considered as resectable, the optimal treatment modality was a combination of radiotherapy and surgery.

Carcinoma, Squamous Cell↗

Structural changes in the round window membrane following exposure to Escherichia coli lipopolysaccharide and hydrocortisone.

The present study focused on structural changes of the round window membrane (RWM) from agents that evoke transient or permanent impairment of the auditory brainstem response when applied into the RW niche. Escherichia coli (E. coli) lipopolysaccharide (LPS) in sterile water (SW) and a 2% suspension of hydrocortisone (CORT), micronized in SW, were instilled into the round window (RW) niche of Sprague-Dawley rats. The morphology of the RWM was analyzed 3 to 21 days after instillation of either substance. Both substances caused minor structural alterations at the light microscopic level. The RWM showed a slight thickening and an invasion of inflammatory cells. At the ultrastructural level, the CORT-treated specimens showed an increased epithelial height and numerous microvilli, whereas the epithelium of the LPS-treated specimens was extended and contained few microvilli resembling those in the normal RWM. We postulate that the RWM may undergo dynamic structural changes when exposed to various agents. The structural alterations per se can influence the passage of substances from the middle ear to the inner ear.

Animals↗

Methods for cellular and subcellular visualization of intermediate filament proteins in the human inner ear.

The preservation of antigenicity for monoclonal antibodies (mAbs) directed against the five classes of intermediate filament proteins and their subgroups was analysed in inner ear specimens from both early embryonic (6-8 gestation-week-old) human labyrinths and inner ears from newborn CBA/CBA mice. After initial fixation in 2% paraformaldehyde, the specimens were embedded in either polyvinyl alcohol (PVA) or the low viscosity acrylic resin LR White. Both embedding media allowed sectioning at room temperature with a specimen thickness of 0.5-1 microns, which gives a resolution at the subcellular level in the light microscope. Immunoreactivity occurred in the PVA-embedded material, but not in specimens embedded in LR White. However, considerably fewer mAbs showed immunostaining in the PVA-embedded material than in both cryofixed-cryosectioned or paraformaldehyde-fixed-cryosectioned human inner ears. Immunoelectron microscopy using colloidal gold (particles 10 nm in diameter) was successful in the PVA-embedded (but not the LR White-embedded) material.

Acrylic Resins↗

Cytoskeletal network of intermediate filament proteins in the adult human vestibular labyrinth.

The cytoskeleton of adult human vestibular hair cells lacks intermediate filament (IF) proteins, whereas in fetal material immunoreactivity for cytokeratin (cks; a subclass of IFs) occurs in both types of hair cells. The shift in the cytoskeletal composition can be hypothesized to their onset of physiological function. Since the IFs are extremely rigid intracellular structures, they provide considerable mechanical stability. The entire cytoplasm of all supporting cells in the epithelial lining of all five vestibular organs is filled with cks. In this way most vestibular hair cells become compartmentalized, each with a rigid shell surrounding it. The distinct delineation of IF proteins in adult tissues, in contrast to fetal inner ear organs in which often a rather general cytoplasmic expression occurs, probably reflects their anatomical basis for their function.

Ear, Inner↗

A low temperature vacuum embedding procedure for X-ray microanalysis of biological specimens at subcellular level.

The validity of freeze-drying and low temperature embedding in Lowicryl resins has been investigated in studies of ion distribution in mouse embryological inner ear, rat rib growth plate, liver and submandibular gland. The morphological preservation of the tissues was adequate for the identification of different intracellular compartments and extracellular structures. It was also possible to analyze extracellular fluids in the vestibular part of the developing inner ear. Compared with thin cryosections, Lowicryl sections are easier to produce and are more stable during analysis. Freeze-dried embedded material can be easily reorientated during cutting and adjacent sections can be used for other purposes such as histochemical and morphological investigations. We found that hydrophilic Lowicryl K11M, which is normally used for immunocytochemical investigations, also can be used for microanalysis. That opens the possibility for combined immunocytochemical and microanalytical studies. However, infiltration and polymerization steps have to be carried out at slightly higher temperatures than when the hydrophobic Lowicryl HM23 is used.

Acrylic Resins↗

The effects of auditory deprivation on morphological maturation of the ventral cochlear nucleus.

The volumes of the auditory brainstem nuclei and age-related auditory brainstem response (ABR) thresholds were analyzed in homozygote (je/je) and heterozygote (je/+) jerker mutant mice. Altogether 97 mice were used in the study. Je/je mice never develop any hearing. The dorsal (DCN) and ventral (VCN) cochlear nuclei were found to have stopped their growth at 56 days after birth. In je/+ mutants, ABR thresholds remained normal or near-normal for 3-6 months, whereas VCN and DCN volumes remained unchanged at least after 56 days after birth. There is no significant difference in DCN volume in je/je and je/+ mice. However, the VCN volume and the cross-sectional area of globular cells were both significantly larger in je/+ than in je/je mice (P less than 0.01). These findings show that auditory deprivation during the maturation of hearing in je/je mutants causes an incomplete maturation of only the ventral cochlear nucleus.

Animals↗

Hydrocortisone applied into the round window niche causes electrophysiological dysfunction of the inner ear.

A suspension of 2% hydrocortisone, micronized in sterile water, was instilled through a perforation in the tympanic membrane into the round window (RW) niche of 9 healthy rats once a day for 5 consecutive days. Three animals were used as controls, and were exposed to sterile water only instilled into the RW niche. Auditory brainstem response (ABR) thresholds were determined for eight frequencies: 2, 4, 6, 8, 12, 16, 20 and 31.5 kHz. Hydrocortisone caused impaired ABR thresholds in the frequency range of 12-31.5 kHz after 5 days of instillation. The impaired thresholds remained unchanged for 2 months, indicating irreversible electrophysiological changes in the inner ear. No morphological damage could be detected in the cochlea by means of light microscopy or transmission electron microscopy. However, hydrocortisone delayed the healing of the perforated tympanic membrane.

Animals↗

Functional changes in the inner ear in renal insufficiency. An experimental study in the rat.

In acute uremia the auditory brainstem response thresholds in the rat (n = 10) became primarily affected in the low-frequency area. As time elapsed in uremia, even the high-frequency thresholds became impaired. In most animals the latencies for wave 2 at the threshold level increased during the first 48 h concomitant with the impairment of thresholds. After 72 h of uremia, the latencies were shortened as compared with the control values.

Animals↗

Lack of differentiation of the isolated murine statoacoustic ganglion during organ culture.

The isolated 12th gestational day statoacoustic ganglion (SAG) of the CBA/CBA mouse was cultured in vitro for 4-8 days. After 4 days, 25% of the SAG had survived, having nerve fibers growing in a circular pattern around the SAG. A further separation into a vestibular and a cochlear part, which occurs in vivo, was not observed. It is concluded that a trophic interaction between the SAG and target tissues is needed for further differentiation of the SAG.

Animals↗

Localization of the integral membrane glycoprotein synaptophysin and the surface glycoprotein Egp-34 in the embryonic and adult human inner ear.

By using well-defined monoclonal antibodies, the distribution of synaptophysin and the glycoprotein Egp-34, the first of its kind, was analyzed immunohistochemically in the fetal and the adult human inner ear. In fetal labyrinths, a distinct immunoreactivity for synaptophysin occurred in the apical region of both outer and inner hair cells as well as in nerve terminals adjacent to both cochlear and vestibular hair cells. In adult hair cells, immunoreactivity was found throughout the cytoplasm. Synaptophysin may act as an important calcium binding protein in the sensory transduction of hair cells. A selective expression of Egp-34 was found in the area of marginal and intermediate cell infoldings in the stria vascularis. A similar localization pattern was found for a number of membrane transport enzymes. The glycoprotein Egp-34 is probably of importance for the active mechanisms regulating the homeostasis of endolymph.

Adult↗

Frequency-specific auditory brainstem response analysis of young normal, aged normal and aged alcohol-addicted rats.

Normative data for frequency-specific (+/- 100 Hz) auditory brainstem response (ABR) thresholds were determined in 60 normal healthy 3-month-old, 5 normal 22-month-old and 6 alcohol-addicted 22-month-old Sprague Dawley rats. The highest degree of auditory sensitivity was in the range of 12-20 kHz. These frequencies showed significant mean differences between the 3-month-old and the old normal rats. In contrast, alcohol-addicted old rats did not reveal any significant threshold changes as compared with either normal old or 3-month-old rats.

Aging↗

Microprobe analysis of cryofixed, chemically untreated freeze-dried cochlear hair cells.

Elemental (energy-dispersive X-ray) microanalysis was performed of cryofixed, chemically untreated freeze-dried cochlear hair cells of 1-month-old CBA/CBA mice. This technique is very difficult to carry out and time consuming, requiring a large number of specimens to be sectioned, quite at random, in order to obtain absolutely perpendicular section planes. Microprobe determination was performed in different parts of outer and inner hair cells. In addition to the normal intracellular elemental content a high calcium concentration was found at the base of all hair cells. Lower calcium levels were evident in the entire cytoplasm, including the cuticular plate and stereocilia. Technically it was feasible to analyze individual stereocilia.

Animals↗