Biomedical subjects
Publications and source records attributed to M A Hotz.
The otolaryngologist has a central role in the detailed examination for dysphagia with its vast differential diagnosis. The carefully elicited medical history and a deliberately focused physical examination are the most important tools to disclose the basis of the dysphagia. Nonspecific blind treatment of dysphagia is obsolete.
The aim of this study was to compare three methods of detection of apoptotic cells: (1) the method based on elution of low molecular weight DNA from the ethanol fixed cells followed by cell staining with DAPI (diamidino-2-phenylindole) or propidium iodide as the DNA fluorochromes, (2) the method of in situ labeling of DNA strand breaks with biotinylated dUTP, utilizing exogenous terminal deoxyribonucleotide transferase, and (3) the method of analysis of DNA denaturation in situ using acridine orange to differentially stain denatured and double-stranded DNA sections following cell exposure to 0.1 M HCl. Cells of the human promyelocytic HL-60 line, treated in vitro with the DNA topoisomerase I inhibitor camptothecin, which selectively triggers apoptosis of S-phase cells, were chosen as a model. The method based on analysis of changes in DNA denaturability was the most sensitive in terms of detection of the earliest changes in chromatin of cells undergoing apoptosis; the increased sensitivity of DNA to denaturation in S-phase cells was measured as early as 100 min after addition of camptothecin. DNA cleavage, assayed either by the univariate measurement of DNA content following extraction of low molecular weight DNA, or by labeling DNA strand breaks with biotinylated dUTP, was detected in S-phase cells after 120 min incubation with camptothecin. The percentage of apoptotic cells at the late stage of apoptosis, the kinetics of cell transition to apoptosis, and kinetics of the loss of S phase cells were all essentially similar when measured by any method.(ABSTRACT TRUNCATED AT 250 WORDS)
Ototoxic drugs, such as aminoglycosides, affect outer hair cell integrity in the inner ear. Transiently evoked otoacoustic emission (TEOAE) characteristics are related to outer hair cell function and can be expected to reflect the influence of ototoxic agents. Transiently evoked otoacoustic emissions were measured during amikacin sulfate therapy in nine patients. The duration of treatment for individual patients ranged from 9 to 33 days. A reversible decrease of overall TEOAE level, occurring after a treatment period longer than 16 days, was found in the majority of patients. The monitoring of TEOAEs is proposed as a method for early identification and, as a result, prevention of aminoglycoside-induced ototoxicity.
One possible alternative to conventional pure-tone testing for screening and monitoring cochlear changes is the measurement of otoacoustic emissions. The aims of this study were to determine the feasibility of using transiently evoked otoacoustic emission (TEOAE) measurements as an objective field procedure and to compare the sensitivity of the measurements indirectly to pure-tone thresholds. The test groups were 117 male recruits and 30 male career cadets in compulsory military service in Switzerland. Transiently evoked otoacoustic emissions were measured before and at the end of a 17-week training period that included exposure to noise from firearms. Results revealed significant changes in response amplitudes in the frequency range from 2 to 4 kHz, whereas changes in the frequency range from 0.5 to 2 kHz were not significant for either group. The changes in relative amplitude did not exceed 15% when spectra containing the lower frequencies were considered. However, they were always greater than 83% within the higher-frequency range. All mean changes were in the direction expected from cochlear damage. Comparison of TEOAE results with pure-tone thresholds measured for a similar sample of subjects indicated that TEOAE testing may be more sensitive than pure-tone audiometry in detecting early cochlear damage from noise. The testing of TEOAEs is feasible as a screening procedure. It offers objective and repeatable information and is substantially less time consuming than pure-tone audiometry.
Exposure of exponentially growing human promyelocytic of lymphocytic leukemic cells to the putative DNA topoisomerase II inhibitor fostriecin (FST), at a concentration of 1 microM, results in the suppression of their rate of progression through the S and G2 phases of the cell cycle. At concentrations between 5 microM and 0.5 mM, FST triggers endonucleolytic DNA degradation in human promyelocytic leukemia cells, resulting in apoptotic cell death; this effect is not selective for any particular phase of the cell cycle. Little or no apoptotic cell death is observed in lymphocytic leukemic cells at any FST concentration. Because FST, unlike other inhibitors of topoisomerase II, such as teniposide (TN) or amsacrine (m-AMSA), does not stabilize cleavable DNA-topoisomerase complexes, the observed differences between the effects of FST versus TN or m-AMSA on the cell cycle may provide clues regarding the role of such complexes in the kinetic effects of these inhibitors. The present results, therefore, are compared with our earlier data on the effects of TN and m-AMSA on the same cells. The only observed difference is the loss of cell cycle phase-specific triggering of DNA degradation by FST in human promyelocytic leukemia cells, compared to the S phase-specific effects of TN and m-AMSA. Therefore, stabilization of the DNA-topoisomerase cleavable complexes may be essential in the selectivity of cell kill during S phase. However, it appears that the presence of stabilized complexes is not essential to the suppression of cell progression through S or G2 or the induction of apoptotis or necrosis, in general, by topoisomerase II inhibitors.
The present review describes several methods to characterize and differentiate between two different mechanisms of cell death, apoptosis and necrosis. Most of these methods were applied to studies of apoptosis triggered in the human leukemic HL-60 cell line by DNA topoisomerase I or II inhibitors, and in rat thymocytes by either topoisomerase inhibitors or prednisolone. In most cases, apoptosis was selective to cells in a particular phase of the cell cycle: only S-phase HL-60 cells and G0 thymocytes were mainly affected. Necrosis was induced by excessively high concentrations of these drugs. The following cell features were found useful to characterize the mode of cell death: a) Activation of an endonuclease in apoptocic cells resulted in extraction of the low molecular weight DNA following cell permeabilization, which, in turn, led to their decreased stainability with DNA-specific fluorochromes. Measurements of DNA content made it possible to identify apoptotic cells and to recognize the cell cycle phase specificity of the apoptotic process. b) Plasma membrane integrity, which is lost in necrotic but not apoptotic cells, was probed by the exclusion of propidium iodide (PI). The combination of PI followed by Hoechst 33342 proved to be an excellent probe to distinguish live, necrotic, early- and late-apoptotic cells. c) Mitochondrial transmembrane potential, assayed by retention of rhodamine 123 was preserved in apoptotic but not necrotic cells. d) The ATP-dependent lysosomal proton pump, tested by the supravital uptake of acridine orange (AO) was also preserved in apoptotic but not necrotic cells. e) Bivariate analysis of cells stained for DNA and protein revealed markedly diminished protein content in apoptotic cells, most likely due to activation of endogenous proteases. Necrotic cells, having leaky membranes, had minimal protein content. f) Staining of RNA allowed for the discrimination of G0 from G1 cells and thus made it possible to reveal that apoptosis was selective to G0 thymocytes. g) The decrease in forward light scatter, paralleled either by no change (HL-60 cells) or an increase (thymocytes) of right angle scatter, were early changes during apoptosis. h) The sensitivity of DNA in situ to denaturation, was increased in apoptotic and necrotic cells. This feature, probed by staining with AO at low pH, provided a sensitive and early assay to discriminate between live, apoptotic and necrotic cells, and to evaluate the cell cycle phase specificity of these processes. i) The in situ nick translation assay employing labeled triphosphonucleotides can be used to reveal DNA strand breaks, to detect the very early stages of apoptosis.(ABSTRACT TRUNCATED AT 400 WORDS)
The effect of magnetic fields on auditory brainstem responses (ABRs) was examined under two conditions. The first involved recording ABRs before and immediately after magnetic resonance imaging (MRI). ABRs were then recorded as the static magnetic field was increased stepwise to 2T. No significant changes in ABR latencies were noted under either condition. These results indicate that MRI has no measurable effect on the transmission latencies of auditory brainstem neural pathways.
The antitumor drug fostriecin (phosphotrienin, FST) has been reported to exert its cytostatic and cytotoxic effects via inhibition of DNA topoisomerase II. The sensitivity of human lymphocytic leukemic MOLT-4 and promyelocytic HL-60 leukemic cells to a wide range of FST concentrations was studied by analyzing the cell cycle-specific effects and changes in nuclear chromatin induced by this inhibitor. The latter was evaluated by assaying the sensitivity of DNA in situ to acid-induced denaturation cytofluorimetrically, with the use of the metachromatic fluorochrome acridine orange (AO), which differentially stains double-stranded and denatured DNA. The cytostatic effects were observed soon after addition of FST (at concentrations of 1-30 microM for MOLT-4 cultures and 1-5 microM for HL-60 cultures) as a perturbation of cell progression through S and G2 phases of the cell cycle. Cell progression through the cycle was halted at greater than 30 microM FST in MOLT-4 cultures and at greater than 5 microM in HL-60 cultures; the effect was instantaneous and affected all phases of the cycle, so that no changes in the cell cycle distribution were apparent with increasing length of exposure to the drug. Instead, at these high FST concentrations, immediate cytotoxic effects became evident, manifesting either as cell apoptosis or necrosis. Apoptosis was observed only in the case of HL-60 cells, at FST concentrations of 5-100 microM, and was characterized by markedly increased sensitivity of DNA to denaturation combined with a decrease in overall DNA stainability, either with the DNA-specific dye DAPI or with AO, indicative of the activation of endogenous nucleases. Necrotic cell death was observed at FST concentrations of 1 mM and at greater than 30 microM for HL-60 and MOLT-4 cells, respectively: in both cases the overall DNA stainability, with either DAPI or AO, was unchanged compared to the control, but their DNA was very sensitive to denaturation. Interestingly, DNA in G2 and late S phase MOLT-4 cells, which were undergoing necrotic death, was much more sensitive to denaturation than was DNA in G1 cells of this lineage. The data indicate that chromatin changes induced by DNA topoisomerase II inhibitors in cells that undergo apoptotic or necrotic death can be conveniently monitored by the assay of DNA in situ sensitivity to denaturation.
The ototoxic effect of plasma-level-controlled netilmicin and tobramycin was monitored in 20 intensive care patients, using auditory brainstem responses (ABRs). ABRs were recorded at the onset and after 6 days of treatment. Significant shifts in the ABR wave V latency were observed independent of the type of medication received or whether the patients were intubated or not. Shifts were also observed in wave I latencies. These latter changes were only significant for the complete population, probably because this potential is more difficult to elicit and measure than is wave V. The I-V interlatency time was greater for netilmicin than for tobramycin. These results indicate that the cumulative effects of aminoglycoside ototoxicity may be different for the central and peripheral nervous systems, and therefore ABR measurements may be the most accurate way of controlling ototoxicity in intensive care patients.