Optical coherence tomography and confocal laser scanning microscopy as non-invasive tools in the diagnosis of sinonasal inverted papilloma: a pilot study.

Optical coherence tomography (OCT) and confocal laser scanning microscopy (CLSM) are light-based imaging techniques that allow for a visualization of microscopic tissue properties in vivo. Our study was to examine whether they allow for differentiation of inverted papilloma (IP) from nasal polyps (NP). Five cases of IP and NP, respectively, were investigated intraoperatively with OCT and CLSM. Biopsies were taken of the investigated area and were analyzed ex vivo with OCT and CLSM and then underwent HE-staining for standard light microscopy. On OCT images, IP showed the characteristic inverted character of the epithelium, that was thicker with a high degree of variability of thickness compared to the thin and homogenous epithelium of NP. In addition, the characteristic stromal edema of NP could be visualized. On CLSM images, the typical epithelial invaginations of IP appeared as crypts, while in NP the highly organized cylindric epithelium could be visualized. In vivo, OCT acquired images of sufficient quality to visualize these characteristics, while CLSM did not. Our study demonstrates that OCT and CLSM can distinguish IP from NP. Further technical development is required to apply the techniques clinically to guide intranasal biopsies or even to make them dispensable.

[Effect of fenestration of the cochlea on the vibration of the round window membrane]. / Effekt der Promontorialfensterung auf die Schwingung der Rundfenstermembran - experimentelle Studie.

Drilling a promontory window and coupling an FMT into the scala tympani may be a surgical alternative to stapes surgery in obliterative tympanosclerosis. Aim of this experimental study on human temporal bones was to measure changes of the acoustic transfer function from the tympanic membrane to the round window membrane after drilling a promontory window and insertion of a floating mass transducer.Laser vibrometry and acoustic measurements were performed on 11 temporal bone preparations equipped with a microphone attached to the round window. Calibrations were carried out to allow determination of SPLs affecting the cochlea after drilling a promontory window leaving the membranous inner ear intact and after insertion of an FMT into the cavity (with or without slight pressure).Drilling a promontory window does influence the transfer function. Insertion of the FMT with additional slight pressure further changes the transfer function.The presence of a promontory window changes the acoustic transfer function to the round window. Further investigations are needed to correlate the qualitative results with the audiological results after "third window vibroplasty" (inserted floating mass transducer without stimulation).

[Advances in endoscopic diagnosis of dysplasia and carcinoma of the larynx]. / Fortschritte der endoskopischen Diagnostik von Dysplasien und Karzinomen des Larynx.

To improve the preoperative and intraoperative diagnosis of laryngeal cancer and its precursors, various endoscopic imaging techniques have been developed in recent years. These techniques differ markedly in their specific applications and goals. Precisely distinguishing among normal mucosa, dysplasia and invasive carcinoma with these procedures is necessary. Furthermore, the exact identification of tumor margins should be possible. The long-term goal is the development of optical biopsy. Since so far there have only been small studies regarding the evaluation of the presented methods, it is necessary to establish multi-center trials with large sample sizes to accurately estimate the value of these endoscopic imaging techniques.

[Mastoid obliteration with a highly porous bone grafting material in combination with cartilage]. / Mastoidhöhlenobliteration mit einem hochporösen Knochenaufbaumaterial (NanoBone®) und Conchaknorpel.

BACKGROUND: An open mastoid cavity might lead to various problems for the patient. Chronic inflammation of the cavity with secretion, changes in the acoustic behavior, vertigo in restricted situations and an impaired self-cleaning function might affect the patient. For surgical treatment reducing of the size of such cavities have been described. Besides autologous materials such as hydroxyapatite or alloplastic substances as tricalcium phosphate have been previously used. A very slow resorption of these materials with rejection has been described. The new ceramic NanoBone® was fabricated in a sol-gel process at 700 °C depositing unsintered hydroxylapatite in a SiO2 structure. This method provides a nano/microstructure of high porosity of the resulting matrix. MATERIAL AND METHODS: 20 patients were reexamined after an average of 2 years and 5 months after obliteration of the open mastoid cavity with NanoBone®. We compared pre- and postoperative findings in terms of otorrhea, frequency of medical consultation, vertigo and otoscopic findings. In 5 patients, in addition, a postoperative CT scan of the temporal bones was used for evaluation of osteoinduction and osteointegration. RESULTS: After obliteration of the open mastoid cavity with NanoBone ® we observed an uneventfully healing. After surgery we achieved a reduction of vertigo, otorrhea and frequency of medical consultations for the single patient. CONCLUSION: The obliteration of an open mastoid cavity with NanoBone ® is a safe alternative method relative to the surgical techniques with autologous materials.

Matrix change of bone grafting substitute after implantation into guinea pig bulla.

BACKGROUND: Many different surgical techniques have been developed to remove open mastoid cavities. In addition to autologous materials, alloplastic substances have been used. A very slow absorption of these materials and extrusion reactions have been reported. We investigated a newly developed, highly porous bone grafting material to eliminate open mastoid cavities, in an animal model. To characterise the transformation process, the early tissue reactions were studied in relation to the matrix transformation of the bone material. MATERIAL AND METHODS: NanoBone (NB), a highly porous bone grafting material based on calcium phosphate and silica, was filled into the open bullae from 20 guinea pigs. The bullae were examined histologically. Energy dispersive X-ray spectroscopy (EDX) was used to investigate the change in the elemental composition at different sampling times. The surface topography of the sections was examined by electron microscopy. RESULTS: After 1 week, periodic acid-Schiffs (PAS) staining demonstrated accumulation of glycogen and proteins, particularly in the border area of the NB particles. After 2 weeks, the particles were evenly coloured after PAS staining. EDX analysis showed a rapid absorption of the silica in the bone grafting material. CONCLUSIONS: NanoBone showed a rapid matrix change after implantation in the bullae of guinea pigs. The absorption of the silica matrix and replacement by PAS-positive substances like glycoproteins and mucopolysaccharides seems to play a decisive role in the degradation processes of NB. This is associated with the good osteoinductive properties of the material.

[Tonsil protrusion in the case of an infraauricular mass. A rare iceberg tumor]. / Tonsillendifferenz bei infraaurikulärer Raumforderung. Ein seltener Eisbergtumor.

We report on a very rare case of iceberg tumor. A giant lipoma arising from the deep lobe of the parotid gland led to an infraauricular mass and tonsil protrusion. After appropriate imaging (ultrasound and MRI) surgical resection was performed. Tonsil protrusions require differential diagnostic evaluation. Apart from the most prevalent parotid gland tumors, like pleomorphic adenoma and cystadenolymphoma, one must consider rare lipomas. Malignancy (e.g. liposarcoma) can only be excluded by histological examination of the whole specimen.

[Optotensometry for measuring the tube function - a feasibility study on a middle ear model]. / Optotensometrie Langzeitmessung der Tubenfunktion - erste Versuche am Mittelohrmodell.

BACKGROUND: The amount of pressure in the middle ear depends mainly on the function of the Eustachian tube. Currently there are no continuous recording techniques measuring Eustachian tube function in clinical context and under physiological conditions over extended periods of time. In this paper we investigate the suitability of an active optic triangulation method on the basis of a projected laser-point-pattern in measuring tympanic membrane movement during pressure variations in a middle ear model. MATERIAL AND METHODS: For projection we used a green semiconductor laser with an output of 1 mW and a diffractive optical element (DOE). As our measured object we used purple latex-foil (Kimberley-Clark (®)), fixed airtight on the cut-off end of a 2 ml syringe-tube. The movement of the foils was measured by an active optic triangulation method. To simulate pathological variations of the tympanic membrane we prepared the latex-foils in specific ways. One foil was perforated and then covered again (simulating tympanic membrane perforation), another one was partly strengthened by sticking a piece of thick, hard paper to it from the inside (simulating calcification). RESULTS: The test-setup, as well as the appliance of pressure-changes worked fine and measurement of foil movement in all the modified foil surfaces was possible. This shows that it is possible to record foil-movement with this system even in tympanic membranes with pathological variations. CONCLUSIONS: In the course of this study we were able to show that it is possible to assess and record foil movement using a system of optic triangulation and to simulate different tympanic membrane pathologies. This could be used both in ENT medicine, as well as in aviation and diving medicine.

[Tensometry - development of a measurement technique of the tube function in a temporal bone model]. / Tensometrie - Entwicklung eines Langzeitmessverfahrens der Tubenfunktion am Felsenbeinpräparat.

OBJECTIVE: Middle ear pressure depends widely on the function of the eustachian tube. Tube dysfunction is often a trigger for middle ear diseases like chronic otitis media but also for barotrauma. Patients with impaired tubal function should not be exposed to situations with extreme pressure changes. Until today, there is no valid examining method for long-time measuring of the development of middle ear pressure. MATERIAL AND METHODS: The basic idea was to develop a thin flexible film with integrated strain gauges made of platinum and gold, which can be applied directly on the surface of the tympanic membrane. Using these, shifts or distensions of the tympanic membrane in a temporal bone model can be measured and documented. RESULTS: We were able to show that the measuring system was sensitive enough to register pressure variations in the middle ear volume of a temporal bone model. While using a full bridge design of the strain gauge resistors it could be shown that the measuring voltage of the strain gauges were in phase of the pressure measurement curve or according to the positioning in opposite phase. In single resistor mode the measurement were not so positioning depended. Measuring tympanic membrane movement in case of perforation was feasible. CONCLUSIONS: To improve the longtime stability of the strain gauge film the next development step will be to make a new design with Konstantan resistors (CU-Ni-Mn). After a testing period, longtime measurements in clinical studies will follow.

Oral pain perception and taste in burning mouth syndrome.

BACKGROUND: Aim of this study was to investigate the effect of idiopathic burning mouth syndrome (BMS) on both, the pain perception within trigeminal lingual nerve distribution and gustatory sensitivity using capsaicin threshold test, and regional taste tests, respectively. METHODS: Pain thresholds for capsaicin were assessed using capsaicin-impregnated filter-paper strips. The strips were placed midline on the tongue tip for whole mouth testing with the mouth closed, and on the left or right edge of the extended anterior tongue for lateralized testing. Measures of gustatory function were obtained by validated "taste strips" test kit and electrogustometry. The tests were applied to 13 patients with BMS. Results were compared with those from 28 healthy subjects. RESULTS: Patients with BMS exhibited a decreased gustatory and somatosensory perception compared with healthy controls. These changes were found for lateralized tests but not for the whole mouth test procedure. Duration of disorder showed an effect on the capsaicin threshold, with patients being less sensitive to capsaicin exhibiting an increased duration of disorder. CONCLUSION: Both pain-related and gustatory sensitivies of the tongue are found to be decreased in BMS.

[Taste disorders. An update]. / Schmeckstörungen. Ein Update.

Clinically, taste disorders are less frequent than smell disorders. Nevertheless, they are very distressing to the concerned patients. Unfortunately, the underlying causes are not always found and a majority of patients are considered idiopathic dysgeusia cases. Besides this taste disorder of unknown origin, medication side effects and surgical injuries are the most frequent origins of dysgeusia. Despite recent, impressive molecular biological and physiological insight into the taste system, few clinical improvements have been achieved. Consequently, curative treatment options remain sparse.

[Devitalization of cholesteatoma on human ossicles by hydrostatic high pressure treatment]. / Devitalisierung von Cholestatomanteilen humaner Ossikel durch hydrostatische Hochdrucktechnik.

BACKGROUND: In cholesteatoma surgery the use of autogenous ossicles for restoration of sound conduction is often limited because of ingrowth of matrix epithelia into the ossicular bone. In an attempt to eliminated these epithelial cells we tested extracorporal high-pressure hydrostatic treatment as a new method for devitalizing the bone but maintaining its structure. METHODS: The inpact of different high hydrostatic pressures either on single cell suspensions of fibroblasts and osteoblasts, or on cells in ex-vivo ossicles chain was examined with different methods (quantitative proceedings, live/dead assay). Additional electron microscopic investigations illustrate the influence of high hydrostatic pressure treatment on cell suspensions of osteoblasts. RESULTS: High hydrostatic pressure between 150 MPa and 250 MPa showed no effect to cellular material. A safe elimination of cell growth was found after an application of pressures at or above 400 MPa. The electron microscopic investigations illustrate clearly the destruction of cellular membranes after high hydrostatic pressure treatment. CONCLUSIONS: These findings give hope that after extracorporal high hydrostatic pressure therapy autogenious ossicles might be used for middle ear recontructions even if they had contact with the cholesteatoma matrix or even were infitrated by keratinized squamous cell epithelia.

[Thermographic investigations during stapedotomy using a CO2 laser]. / Thermografische Untersuchung während der CO2-Laserstapedotomie.

BACKGROUND: Temporary sensorineural hearing losses after CO(2) assisted stapes surgery have been described in literature. Whether thermal effects are the reason for this side effect is still unclear. The specific aim of this research is to demonstrate the thermal spread during CO(2) laser stapedotomy using a high-speed infrared camera. MATERIAL AND METHODS: Thermal depth effects in physiological saline solutions were examined in cochlea models (glass capillary tube, crystal cuvette and temporal bone preparation). Temperature changes and exchange processes after CO(2) laser application were measured using a high-speed infrared camera. Visualization of gas bubbles triggered by vaporisation underneath the stapes footplate after CO(2) shot was achieved with a high-speed camera. The footplate perforations were performed with a scanner system using a micromanipulator (laser power 20 W, 0.6 mm diameter, continuous wave, duration of 40 ms). RESULTS: Homogenous temperature increases of more than 100°C were measured on the surface of the footplate and 0.25 mm within the capillary tube after single shot CO(2) laser stapedotomy. Gas bubbles may cause isolated temperature increases of up to 65 K. These effects were found in depths of up to 2 mm underneath the stapes footplate. CO(2) shots on the saline solutions with parameter used for initial laser application may lead to temperature increases between 20-65 K in depths of 3-6 mm. CONCLUSIONS: CO(2) applications to an opened inner ear with the same laser parameter used for initial shot on the stapes footplate should be avoided.

[Optical coherence tomography in middle ear surgery]. / Optische Kohärenztomographie in der Mittelohrchirurgie.

The application of optical coherence tomography (OCT) in middle ear surgery has a high diagnostic potential, especially for intraoperative evaluation of the cause of stapes fixation, intraoperative assessment of the morphology of the stapes footplate in revision stapes surgery, and as an orientation guide in cochlear implantation in congenital anomalies. OCT displays the middle and inner ear structures precisely. This technology enables the surgeon to use this information for further specification of the intraoperative modus operandi. Considering our own in vivo and ex vivo investigations and animal experiments, potential areas of application can be defined: visualization of the oval window niche in revision stapesplasty and reconstructive middle ear surgery, as well as during explorative tympanotomy for intraoperative assessment of perilymph fistula, and demonstration of structures of the exposed but not opened inner ear.

Habitat continuity effects on gradients of fish biomass across marine protected area boundaries.

Marine protected areas (MPAs) could be useful as fisheries management tools for the exportation of pelagic eggs, larvae and adult fish. A decreasing gradient of fish biomass across MPAs boundary may indicate export. We determine whether gradients of decreasing biomass of fish assemblage occurred in Tabarca Marine Reserve over two habitats with different continuity across the boundaries, to test if the patchy nature of the marine environment might act as a barrier for the fish export. In general, significant decreasing gradients in total fish biomass and biomass of some species were observed on P. oceanica and rocky substrates, independently of their different continuity through the reserve boundaries. Changes in the multivariate structure of the fish assemblage were correlated with the distance from integral reserve. All of these results support the hypothesis that the exportation of adult fish from Tabarca Marine Reserve occurs, and this process may influence the surrounding fished areas.

Temporal bone investigations on landmarks for conventional or endosteal insertion of cochlear electrodes.

CONCLUSION: Our anatomical findings place special emphasis on the requirement to follow an infero-anterior approach to the round window, to expose the scala tympani safely for 'normal' cochlear implantation. It is also known how easily the basilar membrane may be accidentally damaged, despite exercising considerable caution in the approach used. With regard to an 'endosteal electrode' it can be stated that there are no really specific indicators to locate the spiral ligament, or each of the scalae, on the lateral aspect of the tissue layer encasing the cochlea. For the concept of an endosteal electrode, however, the soft tissue layer of the lateral aspect of the cochlea is considered to be sufficiently thick to serve as a physical barrier between the electrode and the inner ear fluid. OBJECTIVES: To re-evaluate surgical techniques of gaining access to the scala tympani for cochlear implantation (cochleostomy, 'fenestration'). There are two reasons for this study. First, recent publications show that in a significant number of patients the electrode array was unintentionally inserted into the 'wrong' scala (sc. vestibuli). Second, dealing with an alternative concept proposed by Lehnhardt for patients with residual hearing ('endosteal electrode'), the anatomical site of the spiral ligament should be known. In a study on human temporal bones the topography of the middle and inner ear is revised with regard to the presence of anatomical or surgical landmarks that may guide the surgeon. MATERIALS AND METHODS: Anatomical examinations were performed on 10 temporal bones (5 fresh specimens and 5 fixed in formalin), in which the bone of the promontory was carefully milled. The consistency of identification and the relative location of specific surgical indicators or landmarks such as 'blue lines' and 'gray lines' were evaluated for 10 temporal bones. Furthermore, the projection of the lateral attachment of the basilar membrane on the promontory was determined with regard to round window anatomy. RESULTS: In all cases, a major blue line indicated the lateral aspect of the basal cochlear turn while milling the promontorial bone. In a limited number of cases (20%), an additional gray line potentially indicated the spiral ligament before the last shell of bone was removed. In 80% of the cases it was possible to remove the bony layer and leave the endosteum intact as a precondition for a potential endosteal electrode insertion. In addition, through the examination of these models, the relative anatomical location of structures, such as the scala vestibuli, scala tympani, spiral ligament, and basilar membrane, is reviewed.

Hydroxyl radical-induced apoptosis in human tumor cells is associated with telomere shortening but not telomerase inhibition and caspase activation.

Reactive oxygen species (ROS) have been found to trigger apoptosis in tumor cells. At the same time, telomerase is found to be associated with malignancy and reduced apoptosis. However little is known about the linkage between ROS such as *OH and telomerase/telomere. To address the interrelations between *OH and telomerase/telomere in tumor cell killing, HeLa, 293 and MW451 cells were induced to undergo apoptosis with *OH radicals generated via Fe(2+)-mediated Fenton reactions (0.1 mM FeSO(4) plus 0.3-0.9 mM H2O2) and telomerase activity, telomere length were measured during apoptosis. We found that during *OH-induced apoptosis, telomere shortening took place while no changes in telomerase activity were observed. Our results suggest that *OH-induced telomere shortening is not through telomerase inhibition but possibly a direct effect of *OH on telomeres themselves indicating that telomere shortening but not telomerase inhibition is the primary event during *OH-induced apoptosis. Strikingly, we also found that *OH-induced apoptosis in HeLa cells is caspase-3-independent but is associated with reduction of mitochondrial transmembrane potential. Our results indicate that *OH triggers apoptotic tumor cell death through a telomere-related, caspase-independent pathway.

Expression of telomerase inhibits hydroxyl radical-induced apoptosis in normal telomerase negative human lung fibroblasts.

In tumor cells telomerase activity is associated with resistance to apoptosis and the introduction of the human telomerase reverse transcriptase (hTERT) subunit into normal human cells is associated with life span extension of the cells. To determine the role of telomerase in regulating apoptosis, telomerase negative human embryo lung fibroblasts were transfected with the hTERT gene. Unlike the control fibroblasts, the telomerase-expressing cells had elongated telomeres and were resistant to apoptosis induced by hydroxyl radicals. The results indicate that expression of telomerase and, thus, the maintenance of telomere length in normal human somatic cells caused resistance to not only cellular senescence but also apoptosis. Moreover, we found that hydroxyl radical-induced apoptosis in telomerase-expressing and control fibroblasts was caspase-3 independent. These findings have revealed a new type of interrelation between telomerase and caspase-3, which may indicate that in this case the expressed telomerase may inhibit apoptosis at a site not related to the caspase-3 cascade.