Wolfgang Arnold

Targeting cancer cells: magnetic nanoparticles as drug carriers

Magnetic drug targeting employing nanoparticles as carriers is a promising cancer treatment avoiding side effects of conventional chemotherapy. We used iron oxide nanoparticles covered by starch derivatives with phosphate groups which bound mitoxantrone as chemotherapeutikum. In this letter we show that a strong magnetic field gradient at the tumour location accumulates the nanoparticles. Electron microscope investigations show that the ferrofluids can be enriched in tumour tissue and tumour cells.

Atu027, a Liposomal Small Interfering RNA Formulation Targeting Protein Kinase N3, Inhibits Cancer Progression

We have previously described a small interfering RNA (siRNA) delivery system (AtuPLEX) for RNA interference (RNAi) in the vasculature of mice. Here we report preclinical data for Atu027, a siRNA-lipoplex directed against protein kinase N3 (PKN3), currently under development for the treatment of advanced solid cancer. In vitro studies revealed that Atu027-mediated inhibition of PKN3 function in primary endothelial cells impaired tube formation on extracellular matrix and cell migration, but is not essential for proliferation. Systemic administration of Atu027 by repeated bolus injections or infusions in mice, rats, and nonhuman primates results in specific, RNAi-mediated silencing of PKN3 expression. We show the efficacy of Atu027 in orthotopic mouse models for prostate and pancreatic cancers with significant inhibition of tumor growth and lymph node metastasis formation. The tumor vasculature of Atu027-treated animals showed a specific reduction in lymph vessel density but no significant changes in microvascular density.

The level and growth behavior of the 2 f1−f2 distortion product otoacoustic emission and its relationship to auditory sensitivity in normal hearing and cochlear hearing loss

The 2 f1-f2 distortion product otoacoustic emission (DP) was measured in 20 normal hearing subjects and 15 patients with moderate cochlear hearing loss and compared to the pure-tone hearing threshold, measured with the same probe system at the f2 frequencies. DPs were elicited over a wide primary tone level range between L2 = 20 and 65 dB SPL. With decreasing L2, the L1-L2 primary tone level difference was continuously increased according to L1 = 0.4L2 + 39 dB, to account for differences of the primary tone responses at the f2 place. Above 1.5 kHz, DPs were measurable with that paradigm on average within 10 dB of the average hearing threshold in both subject groups. The growth of the DP was compressive in normal hearing subjects, with strong saturation at moderate primary tone levels. In cases of cochlear impairment, reductions of the DP level were greatest at lowest, but smallest at highest stimulus levels, such that the growth of the DP became linearized. The correlation of the DP level to the hearing threshold was found to depend on the stimulus level. Maximal correlations were found in impaired ears at moderate primary tone levels around L2 = 45 dB SPL, but at lowest stimulus levels in normal hearing (L2 = 25 dB SPL). At these levels, 17/20 impaired ears and 14/15 normally hearing ears showed statistically significant correlations. It is concluded that for a clinical application and prediction of the hearing threshold, DPs should be measured not only at high, but also at lower primary tone levels.

Suppression tuning characteristics of the 2 f1−f2 distortion‐product otoacoustic emission in humans

The suppression tuning properties of the 2 f1-f2 distortion-product otoacoustic emission (DPOAE) were measured in 16 ears of normally hearing human subjects. DPOAE were elicited by fixed, low-level primary tones in four frequency regions with the second primary tone frequency f2 at 1, 2, 4, and 6 kHz. For various suppressor frequencies, suppression of the DPOAE was measured as a function of the suppressor tone level, enabling the assessment of the threshold and the growth of suppression. Depending on the distance of the suppressor tone to f2, there were marked differences in the suppression behavior of different suppressor frequencies. The threshold of suppression was minimal slightly above f2 and hardly increased with increasing frequency, but increased continuously with decreasing suppressor frequency. The growth of suppression, however, did not systematically change below f2, but decreased rapidly above f2. Both changes resulted in asymmetrical, V-shaped suppression tuning curves. They were sharply tuned to a frequency slightly above f2, with Q10 dB values up to 7.87. This is consistent with the assumption that the main source of the DPOAE is at the f2 site. In some cases, the DPOAE was particularly sensitive to suppressor tones near the DPOAE frequency. In one individual case, facilitation was found for corresponding frequency-level ranges of the suppressor tone. This may suggest a secondary emission source at the distortion product place.

Evidence of Serum Antibodies against Inner Ear Tissues in the Blood of Patients with Certain Sensorineural Hearing Disorders

In healthy human temporal bones, immunoglobulins (IgG, IgA) and the secretory component are demonstrated by two immunohistochemical methods in epithelial cells and in the lumen of the endolymphatic sac. The normal human inner ear does not otherwise show any localization of immunoglobulins. By means of indirect immunofluorescence, antibodies against healthy inner ear tissue were found in the serum of 15 out of 21 patients with double-sided progressive or fluctuating sensorineural hearing loss of undefined etiology. In 2 cases of Cogans syndrome, it was also possible to demonstrate serum antibodies against epithelial structures of the cornea. The lymphocyte mitogen stimulation test performed with the fresh blood of 5 patients with positive detection of serum antibodies against healthy inner ear tissue was negative.

Deep electrode insertion in cochlear implants: apical morphology, electrodes and speech perception results.

Morphological examination of the human temporal bone in the apical region supports the benefits of deep electrode insertion. Initiation of spikes on peripheral processes close to the basilar membrane would provide improved channel selectivity during electrical stimulation but recruiting of nerve fibres requires a higher current. A clinical study was performed on 10 users of the MED-EL COMBI 40 + implant to evaluate the effect of the insertion depth of the cochlear implant electrode on speech perception. All subjects were implanted with the standard COMBI 40 + electrode with an insertion depth of > 30 mm. Acute speech tests were carried out in which stimulation was restricted to the apical, middle and basal regions of the cochlea in turn, and using electrode arrangements in which contacts were either distributed over the whole length of the cochlea or concentrated at the basal end, thus mimicking an insertion depth of approximately 20 mm only. The results showed that stimulation of the apical region of the cochlea supports a significant degree of speech understanding, and that distributing the contacts over the whole length of the cochlea improves speech perception in quiet and in noise.

Growth behavior of the 2 f1−f2 distortion product otoacoustic emission in tinnitus

High-resolution hearing threshold and 2 f1-f2 distortion product otoacoustic emission (DP) were measured with the same in-the-ear sound probe and same calibration at 51 frequencies between 500 and 8000 Hz in 39 sensorineural hearing loss ears associated with tinnitus. Using a primary tone setting L1 = 0.4L2 + 39 that accounts for the nonlinear interaction of the two primary tones at the DP generation site at f2, DPs were elicited in a wide range from L2 = 65 to 20 dB SPL. We failed to find a uniform DP behavior in the 39 tinnitus ears tested. Seventeen of them behaved like impaired ears without tinnitus. In these ears a linearized DP growth was observed where the DP level decreased and the slope of the DP I/O functions steepened with increasing hearing loss and as a result both the DP level and the DP slope strongly correlated with hearing threshold. The other population, 22 tinnitus ears, exhibited a poor or even inverse relationship between DP level and hearing threshold, i.e., displayed an increase of DP level with increasing hearing loss. Despite the severe hearing loss but due to the high level, DPs could be recorded well in the frequency range that corresponded to the appearance of the tinnitus. The DP slope, however, increased with increasing hearing loss and, therefore, did still correlate with hearing threshold revealing pathological alteration. The data suggest that the DP level alone is hardly capable of assessing hearing impairment in tinnitus ears and may even be misleading. Thus just the DP slope seems to be the only reliable indicator of cochlear malfunction around the tinnitus frequency. The observed nonuniform DP behavior suggests different cochlear impairments in tinnitus ears. In those ears where the DP level decreases and the slope of the I/O functions increases with hearing loss, cochlear sensitivity and tuning are supposed to be diminished. In those ears where the DP level increases with increasing hearing loss, a reinforced mechanical distortion is hypothetized to be generated by cochlear hyperactivity that can be the source of both the abnormally high DP level and the tinnitus.

Novel Slow- and Fast-Type Drug Release Round-Window Microimplants for Local Drug Application to the Cochlea: An Experimental Study in Guinea Pigs

Novel drug release microimplants (0.8 × 1.14 mm; custom-made by Leiras, now Schering OY, Finland) of slow- and fast-release types containing either 0.9 mg beclomethasone or no drug at all were placed unilaterally onto the round-window membrane (RWM) of 45 guinea pigs for a maximum duration of 28 days. The following parameters were tested on days 1, 14 and 28 after implantation: threshold levels of beclomethasone in the perilymph of the scala tympani, auditory brain stem responses (ABR thresholds and ABR threshold shifts), RWM morphology and hair cell loss (cytocochleograms). None of the animals in the non-implanted control group (n = 5) or placebo implant group (n = 15), but all animals in the slow-release-type implant group (n = 15) and fast-release-type implant group (n = 15) revealed the presence of beclomethasone on day 1 (34.9 and 64.3 pg/µl, respectively), day 14 (43.8 and 46.9 pg/µl, respectively) and day 28 after implantation (4.7 and 60.5 pg/µl, respectively). Histology of the RWMs appeared normal, and cytocochleograms revealed no inner hair cell loss and outer hair cell loss within normal ranges (from 0.5 ± 0.4 to 0.8 ± 0.2% per cochlea) in both ears in all experimental groups at any time during examination (days 1, 14 and 28). Initial values of ABR thresholds at 3, 6, 9 and 12 kHz did not differ significantly in any of the experimental groups. In non-implanted controls, no significant differences of ABR thresholds were observed in all frequencies tested in either ear on days 1, 14 and 28 compared to initial values, and ABR threshold shifts ranged from –3 ± 5 dB (min.) to +5 ± 7 dB (max.). On day 28 after implantation, there were no significant differences of ABR threshold shifts between this and the implant groups, except for 6 kHz of the slow-release device. Therefore, the placebo implants, the slow-release and the fast-release beclomethasone implants appear suitable for further experiments.

Measles Virus in Otosclerosis and the Specific Immune Response of the Inner Ear

Histologic and immunohistochemical studies of otosclerotic lesions have shown that there is a chronic inflammatory reaction of the otic capsule with bone resorption resulting from vascular invasion accompanied by inflammatory cells. During the active lytic stage of otosclerosis, paramyxoviral structures have been identified by electron microscopy and measles virus antigen expression by immunohistochemistry. Recently, measles virus related sequences have been detected in tissue of otosclerotic lesions. Because the otosclerotic focus has a close relation to the perilymphatic space, the expression of measles virus antigens within it should represent an immunologic challenge to the immune system of the endolymphatic sac. In this study, measles virus specific antibodies were detected in all of the perilymph samples from 19 patients suffering from otosclerosis, and the relative amount of these IgG antibodies was much higher than in serum samples of the same patients or in perilymph of control patients. These findings support the hypothesis that measles viruses play an crucial role in the pathogenesis of otosclerosis.

Diagnostic Procedures for Detection of Lymph Node Metastases in Cancer of the Larynx

Squamous cell carcinoma is the most common malignant neoplasm of the larynx. One of the most important influences on prognosis is the presence of metastases to the cervical lymph nodes. Accurate determination of lymph node involvement is therefore a prerequisite for individualized therapy in patients with squamous cell carcinoma of the larynx. Clinical palpation of the neck is not very accurate and the role of imaging techniques such as ultrasound, ultrasound-guided fine needle aspiration cytology, color Doppler ultrasound, computed tomography, magnetic resonance imaging and positron emission tomography is being applied in order to improve upon the results of clinical investigation alone. According to our investigations and review of the literature, the accuracy of computed tomography scanning (84.9%) and magnetic resonance imaging (85%) was superior to palpation (69.7%) and ultrasound (72.7%). Ultrasound-guided fine needle aspiration cytology showed an accuracy of 89% and was in the same range with positron emission tomography (90.5%).