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Biomedical subjects

M Anniko

Publications and source records attributed to M Anniko.

At least 55 records · Page 3Linked to original sources

Cytokeratin expression patterns by one- and two-dimensional electrophoresis in pars flaccida cholesteatoma and pars tensa cholesteatoma.

Expression patterns of cytokeratins (CKs) in normal skin, in pars flaccida type cholesteatoma (PFTC), and in pars tensa type cholesteatoma (PTTC) were examined by means of one- and two-dimensional electrophoretic techniques. Both CKs 14 and 5 pair (CKs 14/5) and CKs 10/1 were found in all materials. Neither CKs 16/6 nor 19 was found in the skin. CKs 16/6 and 19 were both found in 3 out of 5 PFTCs, only CKs 16/6 in 1 out of 5 and neither CKs 16/6 nor 19 in 1 out of 5. CKs 16/6 and 19 were both found in 1 out of 3 PTTCs, only CKs 16/6 in 1 out of 3 and neither CKs 16/6 nor 19 in 1 out of 3. There was no significant difference in the CKs expression patterns between PFTC and PTTC. The expression of CKs 16/6 and 19 suggested that their matrix epithelia were hyperproliferative. However, not all of the cholesteatomas were always hyperproliferative. Patterns of the terminal differentiation of CKs 1, 5, 10 and 14 in the PFTC or the PTTC were basically the same as those in the skin. In the cholesteatoma, eack CK gradually diminished in molecular weight in the cornified layer and debris. Desmosomal proteins were abundant in skin but not in cholesteatomas.

Adult↗

Cytoskeletal organization of the vestibular supporting cells. Saponin perfusion method for observing intracellular structures by scanning electron microscopy.

The cytoskeletal organization of the guinea pig vestibular supporting cells was investigated employing the saponin perfusion method using scanning electron microscopy. The skeletal framework of a cell was composed of actin, intermediate filaments and microtubules. The membrane bound organelles such as nucleus mitochondria, endoplasmic reticulum, Golgi apparatus, etc. were also well documented. This made it possible to investigate the three-dimensional organization of cytoskeletons as well as their complex interaction with various membrane bound organelles. The intermediate filaments were usually seen surrounding the nucleus and extending through the cytoplasm which connected the nuclear membrane to the plasma membrane, reticular lamina or other cytoorganelles. The intermediate filaments also closely related to the desmosomes. These findings suggest that the intermediate filaments provide mechanical support to the cell and its nucleus. The microtubules were closely related to the secretory granules, Golgi apparatus and intermediate filaments, which supports the idea that the microtubules may control the distribution of intermediate filaments, and also play an important role for the transport of the secretory granules.

Actins↗

Direct puncture of large arteriovenous malformations in head and neck for embolisation and subsequent reconstructive surgery.

Five patients with large arteriovenous malformations (AVM) of the head and neck, which were too large or inconveniently placed for operation alone, were treated by embolisation after direct puncture; two of them were subsequently operated upon. They all recovered without complications. Embolisation of the nidus and subsequent operation is a good alternative for the treatment of large AVM. Ligating the supplying arteries is not a treatment. If the arterial routes to the nidus have previously been closed by ligatures selective catheterisation is impossible, though direct puncture of the nidus is a possibility. The nidus of the AVM can then be obliterated by embolisation either as a treatment, or as a preoperative procedure.

Adult↗

Round window membrane in serous and purulent otitis media. Structural study in the rat.

The rat was used as an animal model to reveal structural alterations in the round window membrane (RWM) during serous otitis media (SOM) and purulent otitis media (POM) over a 6-week period. Comparison of POM animals and control animals showed that the RWM in the former became almost six times as thick as that in controls, whereas that of SOM animals was twice as thick. The structural changes in the RWM in POM animals were confined mainly to the epithelium facing the middle ear cavity and the subepithelial space, which was invaded by inflammatory cells and exhibited dilated vessels. The normal flat epithelium was transformed via cuboidal cells to a cylindric epithelium containing both ciliated and goblet cells. In SOM animals, light microscopy revealed only minor changes in the RWM structure. Ultrastructurally, however, the connective tissue layer exhibited dense aggregations of collagen, increased numbers of fibroblasts, and, in one case, elastic fibers. This last phenomenon was not observed in either POM ears or normal ears. The study showed that various inflammatory conditions of the middle ear, both noninfectious (SOM) and infectious (POM), can cause different structural alterations of the RWM. These structural changes may influence passage through the RWM differently.

Animals↗

Protein patterns in human vestibular ganglion cells and hair cells, with functional interpretations.

Cytoskeletal organization was analysed in the vestibular ganglion cells and in the sensory epithelia of all five vestibular organs of the adult human temporal bone. A pancellular rigidity of the upper surfaces of the vestibular organs, evidenced by large quantities of intermediate filaments and actin, seems of importance for the mechanoelectrical transduction by opposing the motion of the cupulae and statoconial layers. Immunostaining for the calcium-binding protein synaptophysin in the sensory hair bundles and in the area close to the cuticular plates--the first of its kind to be demonstrated in human vestibular organs--indicates the presence of calcium-dependent ionic channels. The hypothesis is presented that the calyx might be involved in a short-loop feed back control of type I hair cells, i.e. of the mechanoelectrical transduction itself. Subpopulations of vestibular ganglion cells were identified by the staining pattern of cytoskeletal proteins, but not by ordinary ultrastructural analysis.

Actins↗

Age-dependent disruption of basal lamina and extracellular matrix formation in L-proline analog treated otic explants.

L-azetidine-2-carboxylic acid, LACA, a naturally occurring vegetable imino acid, can be incorporated into mammal proteins instead of proline. This incorporation has an especially inhibitive effect on collagen secretion. Exposure of embryonic mouse inner ear explants to LACA causes dysmorphogenesis and retarded differentiation, reduces the number of collagen fibrils in the perilymphatic spaces and capsules, and gives rise to a dose-dependent derangement of the basal lamina. In control specimens, both in vivo and in vitro, the inner ear epithelia had a dense contiguous basal lamina overlying a well-developed network of collagen fibrils. When the inner ears were exposed to LACA at a concentration of 150 micrograms per ml of medium, there was a loss of the collagen network and gaps appeared in the basal lamina. At exposure to 300 micrograms LACA/ml, scarcely any collagen fibrils were present and the basal lamina was disrupted in many areas, especially beneath the sensory epithelia.

Animals↗

Cochlear structure and function in a recessive type of genetically induced inner ear degeneration.

An age-related consecutive morphological analysis of the cochlea has been performed in homozygote (je/je) and heterozygote (je/+) jerker mouse mutants. A difference in the time of onset of hair cell pathology was evident between the two, but, when taking place, it showed a similar morphological type of degeneration. The cuticular plate and the stereocilia are particularly vulnerable structures and are the primary sites of damage. The suprastructures on both outer and inner hair cells disintegrate at the same time, irrespective of the level along the basilar membrane. During aging, the je/+ animals showed a progressive age-related impairment of auditory brainstem response thresholds which was correlated semiquantitatively to hair cell pathology.

Age Factors↗

Characteristic ionic composition of endolymph is maintained in cultured inner ear.

Inner ear anlagen from mouse were explanted on the 16th gestational day (gd) and cultured for 5 days, i.e. corresponding to the time of birth. By using energy dispersive X-ray technique an elemental composition characteristic for endolymph was found within the membranous labyrinth of the explants. The sodium to potassium ratio in the endolymphatic space of the cultured inner ears corresponded to endolymph of the 16th gd fetus in vivo. There was no difference in the endolymph compartment between the cochlear and vestibular halves of the in vitro specimens. Differences in Na to K ratio between endolymph of the inner ears and the surrounding medium were statistically significant. Thus, endolymph-regulating mechanisms are active also under organ culture conditions, although not fully optimal.

Animals↗

Genetically induced inner ear degeneration. A structural and functional study.

The jerker mouse mutant has an autosomal recessive mutation, which in homozygotes results in early postnatal degeneration of the sensory epithelia in both the cochlea and vestibulum. An age-related consecutive functional and morphological analysis of the cochlea has been performed in homozygote (je/je) and heterozygote (je/+) jerker mice. Cochlear function was determined with auditory brainstem response (ABR) technique. Heterozygotes were regularly tested to a final age of 12 months. With increasing age they displayed ABR threshold shifts which were of about the same magnitude in the whole frequency range tested (2-31.5 kHz) but extremely variable in individual animals. From homozygote jerker mice no ABR could be elicited. There was a similar morphologic pattern of cochlear degeneration in both homozygote and heterozygote jerker mice affecting primarily the stereocilia and cuticular plate in both outer and inner hair cells. In homozygotes these changes were found in very young animals but in heterozygotes they did not become evident until later in life. All parts of the cochlea became simultaneously affected, that means there was no topographical gradient in the pathological course. The ABR thresholds in 12 months old heterozygotes were correlated semiquantitatively to hair cell pathology in the cochlea. The functional and morphological changes in heterozygotes probably reflect a random influence of the jerker gene vis-à-vis the wild type of gene.

Animals↗

Cochlear effects of hyaluronan applied on ruptured round window membrane.

The purpose of the present experimental study was to determine whether hyaluronan (hyaluronic acid, HYA) exerts any functional or morphologic ototoxic effects on the cochlea when in contact with a ruptured round window membrane (RWM). HYA in 1% solution was applied in the round window (RW) niche of rats (n = 6) prior to perforating the RWM. In a control group (n = 6) the RWM was perforated alone. Cochlear functioning and structure were monitored by recording auditory brainstem responses (ABRs) at 2-31.5 kHz and by scanning electron microscopy. Perforation of the RWM alone resulted in immediate loss of ABR thresholds between 6 and 31.5 kHz in 2 of 6 animals. Similar results were obtained after application of HYA into the RW niche and subsequent RWM perforation. In both treatment groups the mean ABR thresholds and mean latencies for wave II at the ABR threshold returned to the pre-surgical (normal) range after 2 months. Also with respect to the cochlear morphology the results in both treatment groups were alike including minor structural changes in hair cell stereociliae but no loss of hair cells. It is concluded that HYA, when instilled into the middle ear with the inner ear opened, is free from cochlear ototoxic effects. Our present findings enhance the potential usefulness of HYA in otosurgery, even in cases when the inner ear has been opened and/or when one wishes to protect the inner ear.

Animals↗

Repair of chronic tympanic membrane perforations using applications of hyaluronan or rice paper prostheses.

A controlled randomized study was performed in 60 patients with 64 chronic, dry tympanic membrane (TM) perforations. The perforations were randomly allocated to either resection of the perforation rim and instillation of 1% hyaluronan (Healon; HYA) in the perforation gap once daily for 7 days (33 ears) or resection of the perforation margin and application of a sterile rice paper prosthesis (31 ears). The treatment effect was documented by TM photography and morphometric measurements of the perforation area. The hearing was assessed with puretone and high-frequency audiometry. After 2 months, 5 of the HYA-treated perforations (15%) and 4 of the rice-paper-treated TMs (13%) were healed. After 1 year, 18 perforations (9 in each treatment group) were healed. In neither group were there any persistent adverse effects on hearing. It is noteworthy that 28% (18/64) of the chronic, long-standing TM perforations could be repaired by these technically simple and time-saving methods. Both procedures should be considered as easy first-choice alternatives to myringoplasty in selected cases.

Adolescent↗

Acetylcholine receptor localization in human adult cochlear and vestibular hair cells.

The FITC technique using alpha-bungarotoxin visualized the staining pattern of acetylcholine (ACh) receptors in adult human cochlear and vestibular hair cells (HCs) in normal labyrinths and in cochleae with sensorineural hearing loss. Flourescence staining occurred in the cuticular plates of all HCs, indicating that the micromechanics of their suprastructures can act under cholinergic control. Quantitative differences of the fluorescence of ACh receptors occurred between the three rows of outer HCs at the same level in the cochlea and decreasing along a base-to-apex directed gradient. There is strong evidence that the subsurface cisterns are integrated in the efferent nerve system. In the degenerating organ of Corti an uncoupling of the efferent system takes places adjacent to disintegrating HCs, though the staining in the cuticular plates remains until a very late stage in HC disintegration. In vestibular HCs type I, fluorescence is emitted in the supranuclear area of the cytoplasm below the cuticular plate probably indicating an efferent guidance on the afferent nerve transmission directly via the HC itself.

Adult↗

Hyaluronan applied to lesioned round window membrane is free from cochlear ototoxicity.

Hyaluronan (HYA) in 1% solution was instilled into the round window (RW) niche of rats (n = 6) prior to perforating the round window membrane (RWM). Cochlear functioning and structure were then monitored by recording auditory brainstem responses (ABRs) at 2-31.5 kHz and by scanning electron microscopy. Perforation of the RWM alone (n = 6) resulted in immediate loss of ABR thresholds between 6 and 31.5 kHz in 2 of 6 animals. Similar results were obtained after instilling HYA into the RW niche and subsequent RWM perforation (n = 6). After 2 months, ABR thresholds were recorded at all frequencies in the HYA-treated animals, whereas in 2 of the controls no ABR thresholds could be elicited at 20 and 31.5 kHz. However, in both treatment groups the mean ABR thresholds and mean latencies for wave II at the ABR threshold returned to the pre-surgical (normal) range after 2 months. With respect to the cochlear morphology the results in both treatment groups were also alike including minor structural changes in hair cell stereociliae but no loss of hair cells. It is concluded that HYA, when instilled into the middle ear with the inner ear opened, is free from cochlear otoxicity.

Animals↗

Freeze-fracture and immunomorphological analysis of spiral ganglion cells.

Freeze-fracture analysis of adult spiral ganglion cells of CBA/CBA mice revealed two types of membrane specializations. Most cells (type I) had a smooth surface and were surrounded by Schwann cells. Type II spiral ganglion cells showed numerous membrane specializations with well-delineated indentations similar to those previously found on hair cells adjacent to afferent and efferent nerve endings. Immunomorphological analysis (using well-defined monoclonal antibodies directed against different subclasses of intermediate filament proteins) revealed a unique co-expression of neurofilaments, vimentin and cytokeratins in spiral ganglion cells of 8- to 22-week human fetuses.

Animals↗

Regional variations in the expression of cytokeratin proteins in the adult human cochlea.

In the adult human cochlea, a cytokeratin (Ck) network exists along the entire surface of the organ of Corti, enclosing it like a shell. Only the surfaces of the outer and inner hair cells are not integrated in this network. In temporal bone specimens, Ck filaments in Hensen's cells were found to be arranged parallel with and closely apposed to the plasma membrane. In the stria vascularis, Cks were identified only in the marginal cells. Cells in Reissner's membrane and spiral prominence showed varying degrees of immunoreactivity to different monoclonal antibodies directed against Cks. A distinct positivity for Cks was found in most spiral ganglion cells, indicating their presence in all cells. The principal pattern of immunoreactivity was the same in the organ of Corti of the entire cochlea. However, a quantitative gradient in the expression of Cks was observed, with more Cks at the apex than at the base. This was correlated to a difference in the number of Hensen's cells between the two regions. The distinct shell configuration of the Ck network in Corti's organ gives it a tonotopically related difference in rigidity which must be of considerable importance for the perception of sound in the cochlea. The absence of Cks in inner and outer sulcus cells gives them cytoskeletal characteristics of mesenchymal cells with a possible regenerative potential.

Adult↗