Biomedical subjects

F Hafner

Publications and source records attributed to F Hafner.

4 recordsLinked to original sources

PubMed2000

Cytosensor Microphysiometer: technology and recent applications.

The Cytosensor Microphysiometer system detects functional responses from living cells in minutes and offers novel information on cell signalling that is often unobtainable with other assay methods. The principle of the system is based on the measurement of small changes in extracellular acidification, using a light addressable potentiometric sensor (LAPS). Energy metabolism in living cells is tightly coupled to cellular ATP usage, so that any event which perturbs cellular ATP levels--such as receptor activation and initiation of signal transduction--will result in a change in acid excretion. As the extrusion of protons is a very general parameter involved in the activation of nearly all kinds of membrane-bound receptors, receptors can be investigated without prior knowledge of the corresponding signalling pathway. However, by blocking certain signalling pathways inside the cell by means of signal transduction probes, specificity can be brought into the system and the corresponding receptor pathways can easily be elucidated. The aim is to give an overview about Cytosensor Microphysiometer technology and to demonstrate, with the help of some recent applications, the capability of the system to measure acidification rates from a wide variety of cell- and receptor-types coupled to different signal transduction pathways. This feature makes the cytosensor system an ideal tool for acting as a single assay system and circumventing the need for multiple assays.

Adenosine Triphosphate↗

PubMed1999

Modified cryopreservation and xenotransplantation of human parathyroid tissue.

INTRODUCTION: A modified cryopreservation technique for human parathyroid tissue was compared with the standard method using a programmed freezer. METHODS: Total parathyroidectomy was performed in three groups of 6-week-old Rowett nude rats. Group I (control) underwent no transplantation of parathyroid tissue (n=9). After 10 days, the rats of groups II (n=15) and III (n=15) underwent xenotransplantation of 20 mg cryopreserved human parathyroid tissue, which had been stored in liquid nitrogen at -196 degrees C for 1-22 months prior to xenotransplantation. The parathyroid tissue was derived from 15 parathyroidectomized patients with renal hyperparathyroidism. Two tissue samples were obtained from every patient. One sample from every patient was cryopreserved by means of the technique of programmed cryopreservation: programmed freezing of human parathyroid tissue at a controlled rate of -1 degrees C/min to -80 degrees C prior to transfer to liquid nitrogen (group II). The second sample from every patient was cryopreserved by means of a modified cryopreservation technique: immediate placement of human parathyroid tissue in a freezer at -20 degrees C and transfer to liquid nitrogen after 2 h (group III). Calcium and intact parathyroid hormone concentrations in serum were determined over a period of 60 days. Furthermore, individual differences in the calcium concentrations were assessed for each rat on the basis of the calcium levels recorded preoperatively and at day 60. RESULTS: All animals in the control group developed hypocalcemia. Serum calcium concentrations returned to normal levels 60 days after xenotransplantation of parathyroid tissue, which had been cryopreserved using the modified technique (group III) in 12 of 15 rats (80%). At day 60, serum calcium had normalized in 10 of 15 rats (67%) after xenotransplantation of parathyroid tissue cryopreserved using programmed freezing (group II). After modified cryopreservation (group III), the median individual difference in the calcium concentrations was -0.16 mmol/l after programmed freezing (group II). At the end of the study, a median level of human intact parathyroid hormone of 3.5 pg/ml and 2.4 pg/ml was estimated for groups II and III, respectively. There was no statistical significant difference of the individual differences in the calcium concentrations or of the levels of human intact parathyroid hormone between groups II and III. CONCLUSION: Human parathyroid tissue was successfully xenografted in the present experimental study. The results obtained after cryopreservation using the described modified technique were equivalent to those recorded after controlled freezing in a programmed freezer. The simplified cryopreservation technique therefore appears to be suitable for human parathyroid tissue.

Animals↗

PubMed1996

Effect of growth factors on estrogen receptor mediated gene expression.

The proliferation of mammary carcinoma cells can be stimulated by estrogens and various growth factors such as EGF and IGF-I. Steroid hormones and growth factors are understood to exert their effects via different receptors and signal transduction pathways. Recently, it has been shown that growth factors can utilize the unliganded estrogen receptor (ER) as a transcription factor. This study was aimed at identifying the growth factors that can act via the estrogen receptor, and finding new estrogen antagonists that block this activity. Originally, a transcription assay was used in which HeLa cells had been transiently co-transfected with the expression vector for the human ER and a reporter plasmid EREwtc luc. EGF and, to a lesser extent, insulin stimulated the expression of the reporter gene in the absence of estradiol (E2), whereas IGF-I was inactive. The stimulatory effect of E2 and insulin was suppressed when the ER was blocked by the pure antiestrogen ICI 182,780. In ER-positive MCF-7 cells, transfected transiently with the reporter plasmid, EGF had no stimulatory effect on luciferase expression. IGF-I stimulated the transcription to about 50% of the E2 value. Similar activity was found for insulin. The effect of both growth factors was only partly reversed by the addition of a pure antiestrogen. The combination of E2 and IGF-I or insulin led to a synergistic activation of transcription. Because transiently transfected cells do not allow one to study the influence of chromatin structure on gene expression, an MCF-7 subline (MCF-7/2a) was established, in which the reporter construct had been integrated in the genome. IGF-I stimulated luciferase expression in these cells, but showed no overadditive effect with E2. The effects of both agents were completely suppressed by the pure antiestrogen ICI 182,780. These data suggest the existence of an ER-independent mechanism for the activation of the reporter gene in transiently transfected cells, but not in stable transfectants.

Breast Neoplasms↗

PubMed1982

The determination of calcium, glucose, urea and uric acid using the Kodak EKTACHEM multilayer film technology: an evaluation.

Calcium, glucose, urea and uric acid determinations on the "'Kodak Ektachem Four Chemistry Analyzer" were tested for precision and an estimation of the accuracy was performed with our laboratory routine methods using patients' sera and proficiency fluids. Precision -- examined over a period of six weeks -- was very good. Even the very stringent claim of the College of American Pathologists (CAP) -- that analytical dispersion should be less than 1/16th of the normal range -- was fulfilled in almost all control samples. The results from patients' sera measured using both the Ektachem and the routine laboratory methods showed very good correlation. Proficiency fluids tested for calcium, glucose and uric acid all resulted in values well within the assigned interval. Some negative bias was noted for the urea determination however. This could not be completely eliminated even though the samples had been reconstituted with a bicarbonate solution according to the manufacturer's instructions. The linear range of the four determinations was checked: calcium is linear up to 4.2 mmol/l, glucose up to 33 mmol/l, urea up to 60 mmol/l and uric acid up to 1370 mumol/l. Our results confirmed the long-term stability of analyzer and slides to be so high that one calibration per week is sufficient.

Blood Glucose↗