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E Thein

Publications and source records attributed to E Thein.

6 recordsLinked to original sources

Automation of the use of fluorescent microspheres for the determination of blood flow.

Fluorescent-labeled microspheres (FM) are a new tool for the determination of organ blood flow. However, the FM-method is labor intensive, because of the necessity to recover the microspheres from the tissue samples. The aim of this study was to automate the FM-method. A Zymate-Robotic System (Zymark, Idstein, Germany) was modified to handle a novel filtration device. The robot is surrounded by 12 different stations which are necessary to process the samples. It performs the sequential steps which are needed to recover the microspheres from the samples. The dyes are finally released from the FM with a solvent and their fluorescent intensity is measured online using a spectrophotometer (Perkin Elmer, Uberlingen, Germany). The robotic system is able to recover the FM through digestion and filtration of the tissue samples using the new filter, to dissolve the FM and to release the dyes so that their fluorescent intensities can be measured for the calculation of organ blood flow.

Animals

[A new experimental model for repetitive osseous blood supply measurement].

The aim of this study was to establish a novel model permitting repetitive analysis of osseous perfusion over a period of 33 days using the fluorescent microsphere technique. After implantation of two port systems into the right and left carotid artery in New Zealand rabbits (n = 3), fluorescent microspheres were injected into the left ventricle, while blood samples for reference probes were taken from the descending aorta. Using seven different fluorescently labeled microspheres, injections were repeatedly performed starting 3 days after implantation (t = 0) at days 1, 3, 5, 12, 19, 26, and 33. Osseous blood flow was semiquantitatively analyzed by counting the number of trapped microspheres within the bone sections performed through the distal femur condyle (n = 8) using a fluorescence microscope. Over the entire observation period of 33 days, intraindividual variance in the number of trapped microspheres was low while there were marked interindividual differences between animals. The mean osseous perfusion in the three animals evaluated so far remained constant over the observation period of 33 days. The present model is the first to allow repetitive analysis of osseous perfusion over an observation period of 33 days. Using this model, the role of regional osseous perfusion can be studied under conditions such as impaired bone healing following radiotherapy- and/or chemotherapy, implantation of biomaterials, and transplantation of bone.

Animals

Changes in serum osmolarity influence the function of outer hair cells.

Fast motility of outer hair cells (OHC) is thought to be based on a hydromechanic principle. In vitro, the function of OHCs can be disturbed by a change in the osmolarity of the culture medium. Whether changes in the serum osmolarity in vivo can also interfere with OHC motility has not been investigated as yet. Serum osmolarity of New Zealand White rabbits (n = 18) was elevated by a continuous infusion of glucose 40%, decreased by an infusion of aqua dest, or kept constant by an infusion of saline. OHC function was monitored using distortion products of otoacoustic emissions (DPOAE). Input output curves were established between 2 and 5 kHz (geometric mean of f2) with primaries of levels between 35 and 55 dB SPL. Cochlear perfusion was measured using a fluorescence microsphere method. Elevation of the serum osmolarity from 306 +/- 17 mosm/l to 365 +/- 23 induced a decrease of DPOAE between 3 and 12 dB SPL. Cochlear blood flow increased from 0.11 +/- 0.09 to 0.15 +/- 0.10 ml/min/g. When decreasing the serum osmolarity from 303 +/- 9 to 281 +/- 8 mosm/l, only slight changes of the DPOAE could be verified. As in the control group, cochlear perfusion was almost unchanged. In the control group, neither serum osmolarity nor DPOAE changed. Comparable to findings in vitro, increasing the serum osmolarity can lead to a disturbance of OHC function. In patients suffering from sudden hearing loss. dehydration due to physical or mental stress is often observed. This new and promising pathophysiological concept needs further clinical evaluation.

Acoustic Stimulation

A new sample-processing unit for the fluorescent microsphere method.

The use of fluorescent-labeled microspheres (FM) for measurement of regional blood flow is an attractive alternative to the use of radioactive-labeled microspheres. In the FM method the FM have to be completely recovered from the tissue samples in a time- and labor-intensive process. For this reason, a considerable loss of FM is possible. The aim of this study was to develop a filtration device that allows the tissue sample to remain in a single container throughout the procedure to make the process easier and to avoid the loss of FM. The core of the sample-processing unit (SPU) is a single-tube filtration device with a polyamide wire mesh. The protocol for processing tissue from different organs (heart, kidney, liver, spleen, intestine, muscle, bone, lung, brain) was modified and thus shortened significantly. Furthermore, the SPU allows direct filtration of the blood reference sample without previous digestion. Different experiments showed that the SPU in combination with the new protocol excludes the loss of 15-micrometers FM. The modifications of the whole procedure render it faster and highly standardized.

Animals