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Biomedical subjects

W Kuijpers

Publications and source records attributed to W Kuijpers.

At least 19 recordsLinked to original sources

Developmentally-regulated coexpression of vimentin and cytokeratins in the rat inner ear.

In the present study the expression of vimentin-type intermediate filament proteins and cytokeratins was studied immunohistochemically in the rat inner ear from 12 days postconception up to 40 days after birth. With the use of a broad spectrum monoclonal antibody, cytokeratin expression was found to be present in the whole epithelial lining except for the sensory cells, throughout all the developmental stages examined. Vimentin was detected in the mesenchymal cells, the mesenchyme-derived tissues and the intermediate cells of the stria vascularis, confirming their origin from melanocyte precursor cells. In addition, the coexpression of vimentin and cytokeratins in the epithelial lining of the membranous inner ear was found to be developmentally regulated. During the final stages of differentiation, vimentin expression disappeared from the majority of the cell types. In the mature cochlea the coexpression of vimentin and cytokeratins was still found in the supporting cells of the organ of Corti, in the cells of Claudius and in external sulcus cells. As far as we could conclude from this study, the sensory cells showed only vimentin expression but not cytokeratin expression. A possible relationship between vimentin expression in adult epithelial cells of the inner ear and a specialised function of these cells is discussed.

Animals

The early stages of polyp formation.

The purpose of this study is to describe polyp formation in the mucosa of the middle ear as a possible model for the pathogenesis of nasal polyps. In 65 Wistar rats, the eustachian tube was occluded on the left side for up to 20 months; 60 right-sided, nonoccluded ears served as controls. Following occlusion, the middle ear mucosa was examined for signs of polyps. Signs of polyp formation or fully developed polyps were seen in 14 (22%) of the middle ears, and were only seen in middle ears with signs of actual or previous infection. It was established that the first stages of polyp formation include epithelial rupture, proliferation of fibrous tissue through the epithelial defect, and epithelialization of the prolapsed fibrous tissue by proliferation and migration of epithelial cells from the surrounding epithelium until there is full epithelial covering of the polyp. This report discusses whether the described middle ear model may form basis for the explanation of the pathogenesis of nasal polyps.

Animals

Eighth cranial nerve dysfunction in hyperostosis cranialis interna.

Hyperostosis cranialis interna is a recently described autosomal dominant bone disorder characterised by hyperostosis and osteosclerosis confined to the skull, especially the calvarium and the skull base. In the affected family members, we found variable simultaneous involvement of cranial nerves I, II, VII and VIII from late childhood onwards, most likely due to nerve entrapment. Auditory and vestibular functions were followed in 3 young family members for 8 years. At the first examination, pure tone audiograms were normal in all 3 cases and case 1 showed no caloric response in the right ear. During follow-up, this ear developed severe hearing loss progressing to deafness. The left ear showed transient sensorineural hearing loss and a temporarily diminished caloric response. Similar observations were made in case 2. Both cases showed abnormal brain stem auditory-evoked responses during and after the sudden hearing loss, in which initially only wave I was preserved and later on wave V returned with significantly prolonged I-V interval. The latter phenomenon was also observed in case 3 on both sides in the presence of normal audiograms during and after transient unilateral facial nerve paralysis, which was accompanied by bilateral diminished caloric responses.

Adolescent

Expression of cytokeratin polypeptides during development of the rat inner ear.

The expression of cytokeratin polypeptides in the different epithelia of the developing inner ear of the rat from 12 days post conception to 20 days after birth was analysed immunohistochemically, using a panel of monoclonal antibodies. Throughout the development of the complex epithelial lining of the inner ear originating from the otocyst epithelium, only cytokeratins which are typical of simple epithelia were expressed. Cytokeratins 8, 18, and 19 were detectable shortly after the formation of the otocyst from the ectoderm (12 dpc), whereas cytokeratin 7 expression was delayed and first appeared in the vestibular portion and subsequently in the developing cochlear duct. During the development of the different types of specialized cells, differentiation-dependent modulation of the cytokeratin expression patterns was observed. In the mature inner ear, the specialized cell types displayed a function-related cytokeratin expression profile, both in the cochlear and vestibular portion. Cytokeratin expression in the flat epithelium of the vestibular portion suggests a more complex composition of this epithelium than has been established from routine morphology. Remarkably, the cochlear sensory cells were apparently devoid of cytokeratins, but no final conclusion could be drawn on the presence of cytokeratins in the sensory cells of the vestibular portion, because of the difficulty to delineate the cell borders between sensory cells and supporting cells.

Animals

Expression of intermediate filament proteins in the mature inner ear of the rat and guinea pig.

The expression of intermediate filament proteins was studied in the mature inner ear of the rat and guinea pig, using a panel of polyclonal and monoclonal antibodies directed against cytokeratins, desmin, neurofilament proteins and glial fibrillary acidic protein (GFAP). The epithelial lining of the endolymphatic space displayed a complex expression pattern of cytokeratin filament proteins, suggesting greater cell diversity than was known sofar from morphological studies. The cytokeratin antibodies when applied to the inner ear tissues revealed the presence of only cytokeratin polypeptides which are typical of simple epithelia (i.e. nos. 7, 8, 18, and 19). Profound differences in cytokeratin expression patterns were, however, found in the various cell types of both the cochlear and vestibular partition. Remarkably, the sensory cells appeared to be devoid of both cytokeratins and neurofilament proteins. Staining with a 200 kDa neurofilament antibody displayed the presence of different populations of ganglion cells in the spiral ganglion and the vestibular ganglion. There was no reaction with antibodies directed against desmin and GFAP. The great resemblance of the intermediate filament protein expression patterns in the inner ear of the rat and guinea pig indicates a close similarity between the different epitopes.

Animals

The mucosal lining of the Eustachian tube of the rat.

This report deals with a microscopical study on the Eustachian tube of the rat. The tube consists of a small membranous part and a larger part which is supported by cartilage to a varying extent. The epithelial lining is composed of pseudostratified ciliated epithelium and squamous epithelium, both arranged in coherent fields, extending from nasopharynx to the middle ear cavity. The ciliated epithelium, which is continuous with the tracts of ciliated epithelium in the middle ear, provides morphological evidence for the existence of a mucociliary transport system. The part of the tube, that is lined by squamous epithelium is suggested to be involved in facilitating the passage of air. The observations made on the secretory elements refer to a different function of goblet cells and the subepithelial compound glands in Eustachian tube physiology.

Animals

The effect of biphosphonate on induced heterotopic bone.

HEBP (1-hydroxy, ethylidene-1, 1-biphosphonate) inhibited mineralization and was observed in matrix-induced heterotopic bone in rabbits. In one group of rabbits, HEBP was administered continuously until sacrifice 20 weeks after the operation. Another group of animals received HEBP for the first four weeks only. The effect of HEBP on de novo bone formation was determined by histologic and biochemical analyses. Implant alkaline and acid phosphatase levels and implant calcium and phosphate contents were measured. The implants of HEBP-treated animals showed diminished implant resorption and, at the same time, formation of atypical osteoid tissue. Quantitative measurements revealed a decrease of acid phosphatase activity, whereas implant alkaline phosphatase activity was unaffected. The mineralization, as depicted by the implant calcium and phosphate content, was almost completely inhibited during HEBP-administration. These effects were completely reversible after the withdrawal of the drug. Remineralization began directly after discontinuation, and recovered only 12 weeks later. The results of this study confirm reports that HEBP cannot prevent the formation of heterotopic ossification. The only effect would be a delay of mineralization during its administration.

Alkaline Phosphatase

Cytokeratins in induced epidermoid formations and cholesteatoma lesions.

The expression of cytokeratins varies with the type of epithelium, the state of differentiation, and pathological conditions. In this study, the differential expression of cytokeratins in external meatal skin and middle ear epithelium was used for a pathogenetic study of cholesteatoma lesions and infection-induced epidermoid formations in the middle ear of the rat. Immunocytochemistry generally revealed an epidermal-type cytokeratin profile in the cholesteatoma matrix, except for the focal expression of nonepidermal cytokeratins at the invasion front. Comparable observations were made in the middle ear of the rat after an infection-induced invasion of epidermal cells from the meatal skin. An infection-induced-cornifying metaplastic lesion of the middle ear epithelium revealed nonepidermal cytokeratin expression. The results of this combined study suggested that the cholesteatoma specimens studied had an epidermal origin. The expression of nonepidermal cytokeratins was considered to result from a state of hyperproliferation rather than from metaplasia.

Animals

Biocompatibility of a polyether urethane, polypropylene oxide, and a polyether polyester copolymer. A qualitative and quantitative study of three alloplastic tympanic membrane materials in the rat middle ear.

The biocompatibility of porous implants made of Estane 5714 F1 polyether urethane, polypropylene oxide, and a poly(ethylene oxide hydantoin) and poly(tetramethylene terephthalate) segmented polyether polyester copolymer (HPOE/PBT copolymer), which were selected as candidates for an alloplastic tympanic membrane, was assessed after implantation in rat middle ears for periods of up to 1 year. Implantation of the materials led to tissue reactions initially associated with the wound-healing process, whereas after 1 month not only the presence of macrophages and foreign-body giant cells surrounding the implant materials but also implant degradation were characteristic for a foreign-body reaction. Macrophages and foreign-body giant cells dominated the picture of the tissue surrounding polypropylene oxide. The altered morphology of these cells, the persistent infiltration of the implantation sites by exudate cells, and the premature death of five rats in the 1-year group suggest that polypropylene oxide degradation was accompanied by the release of toxic substances. Estane and copolymer degradation did not induce tissue responses reflecting implant toxicity, and tympanic membranes given these alloplasts showed a normal healing pattern. Inclusions in the cytoplasm of macrophages associated with degradation and phagocytosis of all of the polymers under study were found to contain iron, silicon, titanium, and aluminum. Growth of fibrous tissue and bone, the latter into Estane and HPOE/PBT copolymer implants, indicated appropriate implant fixation by tissue, although macrophages and foreign-body giant cells were present as well. Especially the fixation of copolymer by ingrowth of bone seems promising in terms of the amount of bone in the pores and the electron-dense bone/copolymer interface. The latter is indicative for bonding osteogenesis. The HPOE/PBT copolymer is a better candidate for alloplastic tympanic membrane than Estane, and the use of polypropylene oxide cannot be recommended.

Animals

The behavior of alloplastic tympanic membranes in Staphylococcus aureus-induced middle ear infection. I. Quantitative biocompatibility evaluation.

The biocompatibility of dense Silastic implants and porous implants made of Estane 5714 F1 polyether urethane, polypropylene oxide, and an HPOE/PBT segmented polyether polyester copolymer was evaluated during an induced Staphylococcus aureus middle ear infection. The middle ear response to infection seemed not to be affected by the presence of implants made of either of the polymers. Light microscopical morphometry and transmission electron microscopy showed degradation of the porous implants under study, but not of Silastic implants, which were invariably surrounded by a fibrous capsule. This finding, combined with the degree of porous implant degradation, the composition of the tissues surrounding the implants, and the tissue/biomaterial interface reactions are consistent with the results obtained in the noninfected middle ear. Round-cell infiltrates however, were predominantly associated with implants made of polypropylene oxide and HPOE/PBT copolymer; while the presence of (phagocytosed) microbial debris was associated with copolymer. The present findings indicate that with respect to implant behavior in infected surroundings Estane is the best porous material, whereas the behavior of Silastic implants did not deviate from that in non-infected ears.

Animals

Immunohistochemical study of cytokeratin expression in normal and pathologic middle ear mucosa of the rat.

The expression of cytokeratins in the epithelium of the middle ear and external auditory meatus of the rat was studied on cryosections of ethylenediaminetetraacetic acid-decalcified specimens by use of a panel of monoclonal antibodies. The normal middle ear epithelium revealed a complex cytokeratin profile, including regional differences. The induction of sterile middle ear effusions resulted in increased cytokeratin expression. Infective effusions were accompanied by both quantitative and qualitative changes in the cytokeratin expression patterns. The differences observed between the cytokeratin profiles of external meatal skin and those of middle ear epithelium may form a useful tool for research into cholesteatoma development.

Animals

Effect of EDTA on cytokeratin detection in the inner ear.

Immunohistochemical studies on the epithelium of the adult inner ear are difficult to perform without decalcification of the bony capsule. In this study, we examined the effect of decalcifying agents on the immunoreactivity of various cytokeratin antigens in the cochlear duct epithelium of 2-day-old rats, allowing the comparison of fresh and decalcified specimens. Decalcification of unfixed tissue in a solution containing EDTA or EGTA and polyvinylpyrrolidone, at pH 7.4 and 4 degrees C for a maximum period of 2 days, not only preserved the antigen epitopes but even enhanced the staining intensities in comparison with fresh specimens. This enhancement effect, caused by chelating agents and found to be blocked by prior fixation with acetone, is suggested to be caused by unmasking of the antigenic epitopes.

Animals

A device for the production of fresh tumor slices for cytotoxic drug testing.

The potential suitability of thin slices of fresh tumor tissue for the in vitro testing of antitumor drugs was studied. With a simple device, sets of tumor slices of equal thickness were produced rapidly with minimal cellular damage. The sections showed a reproducible stable incorporation of 3H-thymidine for incubation periods of more than 24 h. Preliminary data on the effect of cytotoxic drugs on sections of xenografts of human squamous cell carcinoma revealed a distinct correlation between their in vitro effects on 3H-thymidine incorporation and their in vivo sensitivity in nude mice.

Animals

Cholinergic muscarinic receptors in rat cochlea.

Specific 3H-1-quinuclidinylbenzilate (3H-1-QNB) binding to rat cochlea homogenates occurs to a homogeneous class of binding sites with Kd = 0.13 +/- 0.01 nM and Bmax = 0.57 +/- 0.07 fmol per cochlea. Binding is stereoselectively inhibited by benzetimide enantiomers. Dexetimide was more effective than levetimide in displacing 3H-1-QNB from its binding sites (Ki = 4 x 10(-10) M and 6.5 x 10(-6) M, respectively). Pirenzepine inhibits 3H-1-QNB binding with low affinity (Ki = 2 x 10(-6) M), classifying the binding sites as muscarinic M2 receptors.

Animals

Cochlear and vestibular functions of the rat after obliteration of the endolymphatic sac.

A combined morphological and physiological study on the effect of saccus obliteration on the cochlea and the vestibular labyrinth of the rat is presented. Endolymphatic hydrops was successfully induced in 49% of the animals. It was frequently associated with fistulae of the membranous wall and degenerative changes in the organ of Corti and spiral ganglion. In hydropic ears electrocochleography revealed a CAP threshold increase only in those cases where hydrops was associated with loss of hair cells and ganglion cells. In two of these animals also an increased negative SP was present. Static otolith reflexes were found to be normal in all hydropic ears despite severe lesions of the saccule in some of these. Abnormal canal reflexes measured by electronystagmography and cupulometry were established in animals with hydrops of the semicircular canals and in one animal with a fistula of the saccule.

Animals

Autonomic innervation of the nasal mucosa.

The nasal passages play a crucial role in the protection and functioning of the lower airways. Consequently the nerve supply of the nasal mucosa is extensive, which is related to an immediate and adequate reaction upon a variety of external and internal stimuli. A brief review of the present knowledge on nasal autonomic innervation and pharmacology will be given. There is special attention to more advanced methods, such as radioligand receptor binding techniques, which might augment our insight in the significance of the nervous system in nasal (patho)physiology. Furthermore data on the secretory activity of the nasal glands in the rat and its neural regulation will be accentuated.

Animals

Longitudinal flow of macromolecules in the endolymphatic space of the rat. An autoradiographical study.

Secretion and resorption of sulphated glycoproteins by the epithelial lining of the endolymphatic space of the rat were studied autoradiographically with the use of 35SO4 as a selective marker. This isotope was found to be incorporated into the sensory area of cristae and maculae and a small area of epithelial cells adjacent to the sensory epithelium. Thereafter 35S-labelled glycoproteins were secreted into cupulae and otolithic membranes. No such process could be established in the tectorial membrane. From the gradual disappearance of 35S-labelled glycoproteins from cupulae and otolithic membrane coupled with an accumulation of this substance in the lumen of the endolymphatic sac, the existence of a longitudinal flow under physiological conditions could be concluded. Resorption of glycoproteins by the saccus epithelium was found to be very slow. Accumulation of 35S-labelled glycoproteins in the endolymphatic duct after saccus obstruction indicates that, in addition to the endolymphatic sac, also the duct contributes to the mechanism underlying the longitudinal flow of macromolecules.

Animals