In vitro studies of different irradiation conditions for Photodynamic inactivation of Helicobacter pylori.

Helicobacter pylori (HP) infections are considered to be the main cause for chronic gastritis and gastric ulcers, whereby more than half of the world's population is nowadays infected. The increased use of antibiotics is leading to an enhanced resistance. Photodynamic inactivation of bacteria seems to be a potential alternative for antibiotic therapies. In our study we used the photosensitizer Chlorin e6 (Ce6) in combination with red light-emitting diodes to inactivate HP in vitro. Ce6 uptake is determined by spectroscopy. Furthermore diverse experiments of different concentrations in the range of 0-100 µM of the photosensitizer and exposure times up to 300 s are carried out in order to find optimal irradiation parameters (wavelength: 660 nm, power density: 9 mW/cm(2), absorbed dose: up to 2.7 J/cm(2)). The data show a significant reduction after already a few seconds of illumination, even with a low Ce6 concentration in the sub-µM-region. At a concentration of 100 µM a nearly total inactivation (6-log10-reduction) of HP was achieved within 60s of irradiation.

Pharmacokinetic of ALA and h-ALA induced porphyrins in the models Mycobacterium phlei and Mycobacterium smegmatis.

Photodynamic inactivation (PDI) of bacterial strains presents an attractive potential alternative to antibiotic therapies. Success is dependent on the effective accumulation in bacterial cells of photochemical substances called photosensitizers, which are usually porphyrins. It is also important to know the distribution of the photosensitizer in bacteria at the microscopic level. The present results examine the accumulation of photosensitizers by Mycobacterium phlei and Mycobacterium smegmatis, which serve as models for the important pathogens Mycobacterium tuberculosis, Mycobacterium leprae and Mycobacterium bovis. The kinetics of porphyrin synthesis after treatment with the precursors ALA and h-ALA were studied. The goal was to describe the biosynthesis and the pharmacokinetics of sensitizers in both bacterial strains using fluorescence microscopy and spectroscopy. We could show that both Mycobacterium strains enrich porphyrins after ALA and h-ALA administration detected by fluorescence peaks at about 620nm. By HPLC analyses the major porphyrin could be identified as coproporphyrin. In the future we will apply the new knowledge in in vitro and in vivo experiments to strains of M. tuberculosis, M. leprae and M. bovis and examine cell destruction by PDI.

Autofluorescence detection of tumors in the human lung--spectroscopical measurements in situ, in an in vivo model and in vitro.

To detect bronchial carcinoma by autofluorescence, we measured the spectra of tumor and normal tissue in situ, in an in vivo model and in vitro by fiber optic spectrometer and two-dimensional resolved microspectroscopy. The in situ measurements were performed in bronchi of nine patients with squamous cell carcinoma during regular bronchoscopy with autofluorescence assistance. The fluorescence was monitored with a fiber optical spectrometer under blue light excitation (lambda=405nm). In an in vivo model, the resected lobe of a lung was perfused under physiological conditions. Tumorous and normal tissues were examined spectroscopically during perfusion and after blood removal and substitution with formol. In another setup the wavelength dependency of autofluorescence was examined on resected parts of physiological bronchi and central bronchial carcinomas. Under the variation of the excitation from 385 to 465nm the autofluorescence response was monitored with a fiber optic spectrometer. For investigation of the origin of autofluorescence, two-dimensional resolved spectroscopy was performed with the SpectraCube system on several sections of tumor and normal tissues All measurements, performed in vivo, in the in vivo model and in vitro agreed, that the main difference of the autofluorescence between tumor and normal bronchus tissue is the intensity of the fluorescences' main peak at 505nm. The signal on tumor tissue is in all cases significantly lower than that of normal tissue. The shape of the autofluorescence peaks is in healthy and carcinoma tissue approximately the same with two characteristic minima at 540 and 580nm. After the preparation with formaldehyde those minima disappeared from the spectra. A comparison with the absorption spectra of hemoglobin showed, that the variation of the spectra may be due to the blood content in the tissue. Two-dimensional spatially resolved spectroscopy showed, that the lower intensity of fluorescence in tumor tissue is due to the irregular and low-concentrated formation of fluorescent structures, which seen to be the elastic structures of bronchial tissue.

Ablation of vitreous tissue with erbium:YAG laser.

PURPOSE: Using a noncontact erbium (Er):yttrium--aluminium--garnet (YAG) laser, ablation of vitreous was compared to distilled water in vitro. METHODS: The porcine vitreous body and distilled water were ablated in vitro at different pulse lengths and pulse energies. Selected pulse energies were 25, 35, 45, 75, and 100 mJ (pulse rate: 1 Hz; laser beam diameter at the surface of the sample: 2 mm). Pulse lengths were at 140 +/- 3 microsec, 190 +/- 4 microsec, and 240 +/- 5 microsec. The loss of weight in vitreous tissue and distilled water was measured using precision scales and corrected for evaporation, respectively. The Mann-Whitney U test was used to assess the significance of differences in ablation rates of water and vitreous. P < 0.05 was considered statistically significant. RESULTS: Reproducible and constant ablation rates were found in both vitreous and distilled water in each of 10 consecutive series of 50 laser pulses at constant laser parameters. Ablation rates per pulse (microg/microsec) of vitreous tissue were as follows: 3.0 microg to 45.8 microg (140 microsec), 10.4 microg to 53.8 microg (190 microsec), and 17.9 microg to 24.2 microg (240 microsec). The ablation rates exhibited a linear correlation with increasing pulse energies and also with decreasing pulse lengths. Considering the pulse lengths of 190 microsec and 240 microsec with all pulse energies tested, the ablation rates of distilled water were significantly higher (P < 0.05) than ablation of vitreous tissue. The ablation rates at a pulse length of 140 microsec were not significantly different. The differences per pulse were as follows: 0.5 microg to 2.1 microg (140 microsec), 1.9 microg to 6.0 microg (190 microsec), and 3.5 microg to 8.7 microg (240 microsec). CONCLUSIONS: Vitreous ablation is possible using Er:YAG laser. The ablation characteristics of vitreous have proved to be similar but not equal to that of water.

Photoablation of meningiomas and neurinomas by Holmium Yag laser radiation.

Cranial rat bone was irradiated by 2.1 microns Holmium Yag laser radiation. Quantitative edge rates were calculated. Histologic sections were investigated by light and electron microscopy. Eighteen cases of hard fibrous or calcified spinal and cranial meningiomas and neurinomas were operated upon using pulsed laser beam. In rat cranial bone ablation rate ranged between 0.3-0.5 mm per pulse. Perifocal thermal damage was observed in a zone of 20-90 microns around the lesion. In all human cases tumors could be removed totally without additional neurological deficit. In vivo heat development was measured by an i.r.-camera.

Laser Resistance of Endotracheal Tubes II: ObservedTemperature Rise and Theoretical Explanation.

An infrared camera was used to measure the temperature rise which takes place in endotracheal tubes exposed to a 20 W CO2 laser beam. It was seen that a metallic tube was heated up within 1 s to temperatures of 200-300°C which was very destructive to the PVC conduits inside the tube. A compound tube, on the other hand, reached temperatures of only 60°C at its inner surface after an exposure of 20 s. The experimental results can be explained by a physical model which uses the heat conduction and the heat capacities of both tubes. Whereas heat conduction in the metal tube is isotropic, heat conduction in the compound tube is anisotropic with a high conductivity along the outer surface and a low conductivity to the inside. This anisotropy and the cooling mechanism in the compound tube due to vaporising water are the reason for the high laser resistance of the tube.

Laser resistance of endotracheal tubes I: experimental results of a compound tube in comparison to a metallic tube.

Laser-induced ignition of endotracheal tubes is a serious problem in CO2 laser surgery of the upper aerodigestive tract. A new tube built of composite materials with a foam at its surface is protected by vaporisation of water from the wet foam. This tube and a flexible metallic tube of stainless steel were tested experimentally against the radiation of various lasers such as CO2, holmium/YAG, Nd/YAG, Nd/YAG frequency doubled in KD*P, argon and excimer. At laser power densities of 10(3) W/cm(2), the shaft of the metallic tube was damaged within seconds whereas the compound tube was stable for more than 2 min; the damage threshold of the new tube was found to be at 3.2×10(6) W/cm(2). While the compound tube was found to be sufficiently laser resistant, the metallic tube was found to exhibit functional damage, in particular the PVC inflation conduits inside of the tube were destroyed.

Interaction of holmium laser radiation and cortical bone: ablation and thermal damage in a turbid medium.

The ablation of cortical bone by holmium laser radiation is described by experimental values of the ablation rate, the depth of tissue damage, and the tissue temperature. An ablation model is presented on the basis of photon diffusion in a turbid medium. When this model is compared with experimental results for the ablation rate, the penetration depth is determined. The expansion of the laser-induced heat can be explained by a point heat source located in a distance beneath the surface equal to the ablation depth. The accumulation of heat as a function of the repetition rate of the laser leads to a limitation of the repetition rate. In order to avoid traumatic heat accumulation, a maximum repetition rate should not be exceeded.

Effects of simmer current on flash-lamp impedance and their combined influence on the output of the Ho,Cr,Tm:YAG laser.

The increase in slope efficiency and the reduction of the laser threshold of a flash-lamp-pumped Ho,Cr,Tm:YAG laser that are due to increased flash-lamp efficiency when the current of the dc simmer is increased are described. Investigations of the electrical characteristics of the flash lamp as a function of the simmer current prove that the flash-lamp impedance decreases to a constant value when the dc simmer current is increased. As a consequence an increase of the peak current through the flash lamp is observed, which improves the flash-lamp efficiency.

[Measuring vibrations by laser doppler vibrometry of the human tympanic membrane after stapes fixation]. / Laserdopplervibrometrische Schwingungsmessungen am menschlichen Trommelfell nach Stapesfixierung.

At times preoperative diagnosis of otosclerosis can prove difficult. Even the objective measurement of the stapes reflex is not very conclusive in patients suffering from more severe hearing loss. Laser-Doppler vibrometry measures the velocity or amplitude of the tympanic membrane (umbo) vibrations. Laser-Doppler vibrometry has a sensitivity that is highly by the factor 10(4) than tympanometry. The herewith presented examinations show, that there is a correlation between the frequency patterns of the displacement amplitudes and experimentally simulated otosclerosis.

Middle ear transmission disorders--tympanic membrane vibration analysis by laser-Doppler-vibrometry.

Laser-Doppler-vibrometry is a useful method to measure vibrations of the tympanic membrane. It is suitable to objectively diagnose the middle ear. In comparison to tympanometry, laser-Doppler-vibrometry has greater sensitivity. Temporal bone specimens were specially prepared to simulate middle ear disorders like middle ear effusion, fixation of the malleus head, fixation of the stapes footplate and removal of the incus. A correlation between the displacement function of the umbo and the experimentally produced middle ear disorders could be demonstrated. For the first time it was possible to measure the vibration of the tympanic membrane by using a laser-Doppler-vibrometer in a healthy 25-year-old male.

[Laser Doppler vibrometry of the tympanic membrane. Possibilities for objective middle ear diagnosis]. / Laser-Doppler-Vibrometrie (LDV) des Trommelfells. Möglichkeiten für eine objektive Mittelohrdiagnostik.

In the diagnosis of hearing disorders, the laser velocimeter can be used to advantage. In contrast to previously published studies on measurements at exposed middle and inner ear structures of the human temporal bone, we used a clinically more practical method to obtain results through the intact auditory meatus. We used a laser intensity of less than 1 mW. The measurements were performed in a room with unlimited sound transmission of frequencies between 0.5 and 8 kHz. The signals from the vibration of the tympanic membrane, obtained by this touch-free method, were analyzed by Fourier transform, showing only the fundamental oscillation but not the higher harmonics. Experimental data on middle ear effusions, obtained by fixation of the malleus head and by drilling the temporal bone, are presented.