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W G Eisert

Publications and source records attributed to W G Eisert.

At least 19 recordsLinked to original sources
PubMed1990

Relationship between vessel wall 13-HODE synthesis and vessel wall thrombogenicity following injury: influence of salicylate and dipyridamole treatment.

We performed studies to determine the relationship between injured vessel wall thrombogenicity, vessel wall 13-hydroxyoctadecadienoic acid (13-HODE) synthesis and cAMP levels in rabbit treated with salicylate or dipyridamole. Injured vessel wall thrombogenicity was measured as the number of 3H-adenine labelled platelets adhered to the subendothelial basement membrane exposed by air injury in carotid arteries of rabbits treated orally with salicylate or dipyridamole. Vessel wall 13-HODE was measured by HPLC and vessel wall cAMP was measured by RIA. Vessel wall thrombogenicity was increased two-fold in rabbits treated with salicylate and decreased by half in rabbits treated with dipyridamole. The levels of vessel wall cAMP levels were correlated both with the plasma dipyridamole levels and increases in 13-HODE synthesis. cAMP levels were unaffected by salicylate treatment, but 13-HODE synthesis was decreased. We conclude that there is a significant relationship between vessel wall cAMP levels and 13-HODE synthesis, which in turn, influences subsequent vessel wall thrombogenicity.

Animals↗
PubMed1990

Dipyridamole--evaluation of an established antithrombotic drug in view of modern concepts of blood cell-vessel wall interactions.

The effect of dipyridamole on the local antithrombotic activities of endothelium has been evaluated. Human whole blood was allowed to flow over an endothelial cell-derived extracellular matrix partially covered by human endothelial cells. Half-maximal suppression of platelet aggregate formation occurred with approximately 5 microM dipyridamole. Similarly, a pronounced inhibition of thrombus formation was observed by in vivo microscopy and computer-assisted morphometric analysis, after oral treatment of non-anesthetized hamsters with dipyridamole, 5 mg/kg. This strong suppression of thrombus formation was maintained in animals on a long-term cholesterol-supplemented diet. The antithrombotic potential of dipyridamole has been clearly demonstrated, both in vitro and in vivo using these more complex approaches employing quantitative microscopy.

Animals↗
PubMed1990

Dipyridamole alone or combined with low-dose acetylsalicylic acid inhibits platelet aggregation in human whole blood ex vivo.

1. In a randomized, double-blind trial we compared the inhibition of the platelet-vessel wall interactions in whole blood ex vivo. There were four groups of 24 healthy volunteers each of whom were treated orally for 3.5 days with either 200 mg dipyridamole (sustained release preparation), 25 mg acetylsalicylic acid, both drugs combined or placebo twice daily. 2. The mean area of all platelets/aggregates was reduced by 6.2% +/- 4.2% (+/- s.e. mean) by placebo (n = 23), 19.8% +/- 6.7% by dipyridamole (n = 22), 53.7% +/- 4.9% by acetylsalicylic acid (n = 23) and 71.4% +/- 3.7% by the combination of both drugs (n = 24), when compared with total inhibition of aggregation by EGTA. Thus, low-dose acetylsalicylic acid inhibited aggregation (P less than 0.001). 3. Dipyridamole reduced the size of platelet aggregates (P less than 0.01, two-fold analysis of variance). The reduction was correlated with the individual dipyridamole plasma levels (P less than 0.05, analysis of covariance). The subgroup of large and very large thrombi being formed was also reduced by dipyridamole (P less than 0.05). 4. This ex vivo study demonstrates that dipyridamole alone inhibits formation of thrombi on subendothelial matrix and enhances the inhibitory effect of low dose acetylsalicylic acid in this model of thrombosis.

Adult↗
PubMed1989

Thrombin stimulates inositol phosphate accumulation and prostacyclin synthesis in human endothelial cells from umbilical vein but not from omentum.

We have compared the effects of thrombin on the accumulation of inositol phosphates and the synthesis of prostacyclin in cultured human endothelial cells from umbilical vein and the microvasculature of omentum. Active human thrombin induced a dose-dependent accumulation of inositol phosphates and a concomitant synthesis of prostacyclin in endothelial cells from human umbilical vein. However, thrombin at all concentrations tested was unable to stimulate inositol phosphate accumulation and prostacyclin synthesis in microvascular endothelial cells from human omentum. Bradykinin was able to stimulate these effects in both types of cell. These results demonstrate that although inositol phosphate turnover is an initial event associated with prostacyclin synthesis in endothelial cells, there are differences in the way microvascular endothelial cells respond to thrombin.

6-Ketoprostaglandin F1 alpha↗
PubMed1989

Efficacy of single intracisternal bolus injection of recombinant tissue plasminogen activator to prevent delayed cerebral vasospasm after experimental subarachnoid hemorrhage.

Premature lysis of subarachnoid blood clots by thrombolytic substances such as urokinase and plasmin has been shown to be efficacious in preventing cerebral vasospasm in clinical and experimental investigations. Recently, tissue plasminogen activator (rtPA) derived from recombinant deoxyribonucleic (DNA) technology has been introduced as a new thrombolytic substance. With its high affinity for fibrin-bound plasminogen and low affinity for circulating plasminogen by which a clot-selective fibrinolysis can be achieved without the danger of inducing systemic fibrinogenolysis, rtPA might be the ideal substance for the postoperative lysis of cisternal blood accumulations after subarachnoid hemorrhage. The efficacy of rtPA in preventing delayed cerebral vasospasm after experimental subarachnoid hemorrhage using a single intracisternal bolus injection of this agent was investigated. With a single injection of 25 micrograms of rtPA into the cisterna magna 48 hours after the first and 6 hours after the second injection of blood in the two-hemorrhage model of cerebral vasospasm, angiographic spasm of the basilar artery was completely prevented in all animals so treated whereas in the control group severe vasospasm occurred. Autopsy studies of the experimental animals demonstrated that the subarachnoid blood clots were almost completely removed by intracisternal rtPA application. Additionally the pathomorphological signs of proliferative vasculopathy present in all animals of the control group were not demonstrable in the rtPA group. As intracisternal bolus injection of rtPA is highly efficacious in preventing angiographic as well as pathomorphological vasospasm, it is concluded that use of this thrombolytic substance might be a promising approach for pharmacological blood clot removal.

Animals↗
PubMed1989

Denaturation and condensation of intracellular nucleic acids monitored by fluorescence depolarization of intercalating dyes in individual cells.

The intercalating binding of planar aromatic dye molecules to nucleic acids can be analyzed using fluorescence depolarization measurements of the dye molecules excited by linearly polarized light. In this study, we investigated the conformational changes of the intracellular DNA-dye complex in single cells. Flow cytometry, combined with a newly developed double-beam autocompensation technique, permitted rapid high-precision fluorescence depolarization measurements on a large number of individual cells. The dyes ethidium bromide (EB), propidium iodide (PI), and acridine orange (AO) were used in this study. Depending on the dye-to-phosphate ratio of the nuclear acid-dye complex, as well as on the spatial dye structure itself, internal and external binding sites can be monitored by fluorescence depolarization analysis. Both energy transfer and rotation and vibration of the dye molecules cause depolarization of the fluorescence emission. Differences in the concentration-dependent dye fluorescence depolarization values between PI and EB on one side and AO on the other side can be interpreted as a denaturation and condensation of double-stranded DNA regions by AO. We further show that the fluorescence polarization measurement technique can be used in an alternative way to monitor thermal denaturation of cellular DNA.

Acridine Orange↗
PubMed1987

Two-parameter data acquisition system for rapid slit-scan analysis of mammalian chromosomes.

A data acquisition system is described for recording two independent signals simultaneously from a laser-based flow cytometer for rapid slit-scan chromosome analysis. High-aperture microscope optics allow recording of fluorescence distributions along the longest axis of metaphase chromosomes with a spatial resolution better than 1 micron. Fluorescence and small angle forward light scatter as well as dual-wavelength fluorescence signals from Indian muntjac chromosomes stained with propidium iodide (PI) or acridine orange (AO) have been recorded simultaneously. While maintaining the multi-user operation of the computer, photomultiplier signals are digitized at a rate of 400 signals per second, stored temporarily in high-speed cache memories, and transferred subsequently to a minicomputer for further storage. Extensive software packages for data acquisition, analysis, and display of the results are described. Data acquisition is generally done in list mode, allowing complete reconstruction of individual signals (profiles) at any time. The distribution of stained constituents along the chromosomes can be displayed. Furthermore, histograms of various parameters of the input signals may be generated.

Acridine Orange↗
PubMed1986

Different effects of aspirin, dipyridamole and UD-CG 115 on platelet activation in a model of vascular injury: studies with extracellular matrix covered with endothelial cells.

Cultured endothelial cells produce an extracellular matrix (ECM) which activates platelets, similarly to deendothelialized vascular segments. Platelet-rich plasma (PRP) was incubated with endothelial cells cultures seeded in various densities on ECM. The interaction of the platelets with this artificial intima was evaluated by phase microscopy and by thromboxane A2 (TXA2) and prostacyclin (PGI2) measurement. Large platelet aggregates were formed on exposed ECM. Platelets aggregation but not adhesion on the ECM was markedly inhibited by the presence of endothelial cells. Pretreatment of the endothelial cells with 0.1 mM aspirin reduced their PGI2 synthesis and was associated with platelet aggregation on the ECM. 10 microM dipyridamole markedly inhibited platelet activation by ECM when the drug was added to citrated whole blood before PRP preparation. UD-CG 115 which elevates cyclic AMP in cardiac muscle, inhibited platelet aggregation and TXA2 production induced by ECM, in the presence as well as in the absence of endothelial cells, without any effect on endothelial PGI2 production.

6-Ketoprostaglandin F1 alpha↗
PubMed1986

A pulse generator simulating slit-scan chromosome analysis signals.

A simple circuit is described for generating a variety of electronic pulses to test hardware and software for slit-scan chromosome analysis in a flow cytometer. The pulse shape can be changed to have different numbers of local minima, thereby simulating fluorescence pulses from acrocentric, monocentric, and dicentric chromosomes. Long pulses simulate aggregates of chromosomes. The pulse repetition rate as well as the pulse amplitude is variable. Although the circuitry is built with only three integrated circuits, the pulse-to-pulse variation in shape and height is quite small. After digitization of the analog signals, the constructed histograms of pulse integrals show a relative coefficient of variation below 1%. This signal generator provides a valuable tool for a number of electronic test applications that would otherwise require expensive standard particles analyzed in a well-tuned flow cytometer.

Animals↗
PubMed1986

SCAN: a program library for high resolution slit-scan analysis of chromosomes in flow cytometry.

Computer programs for high resolution slit-scan chromosome analysis in flow cytometry are introduced. A modular program library, SCAN, contains programs for single and dual parameter data acquisition, correction of recorded profiles and computation of histograms of various parameters. Using a minicomputer system, data acquisition is programmed in assembler to realize high input rates and real-time histogram calculation. Software for the processing of recorded profiles has been written in FORTRAN and allows extensions or alterations for different objectives. A sample run recording bicolor fluorescence profiles from metaphase chromosomes demonstrates the main features of the software.

Animals↗
PubMed1985

Double-beam autocompensation for fluorescence polarization measurements in flow cytometry.

The degree of depolarization of fluorescent light emitted from an organic dye, which is used as molecular probe, is a powerful tool in probing the microenvironment. By fluorescence depolarization the macromolecular structure can be investigated as well as the the mobility of the marker molecule itself or of the complex formed by the probe. Additional information such as energy transfer rates, donor-acceptor distances, and orientations are also measurable. These data are of particular interest if they can be measured from whole cells. Using flow cytometry, we can analyze a large number of cells with high statistical significance in a short period of time. We describe a newly developed double-beam epi-illumination arrangement for fluorescence polarization measurements that uses an autocompensation technique. This new technique permits the various depolarizing effects within the optical as well as the electronic components of the system to be continually compensated for on a cell by cell basis. Simultaneous measurements of other cell parameters for cell cycle analysis by total fluorescence intensity remains possible. The sensitivity of the system to measure polarization was determined as +/- 0.006 p (0 less than or equal to p less than or equal to 0.5 in isotropic media), which amounts to +/- 1.2% of the maximum p value. Polarization data for latex microspheres plotted in the histogram mode were measured with a standard deviation of 0.006, which proved the high resolution and the high performance of the system.

Animals↗
PubMed1979

Kinetics of carboxymyoglobin and oxymyoglobin studied by picosecond spectroscopy.

Picosecond studies of carboxymyoglobin (MbCO) and oxymyoglobin (MbO2) reveal that excitation at 530 nm induces photodissociation at less than 8 ps. The kinetic and structural changes were monitored by following absorbance changes at selected wave-lengths in the Soret (B) band and in the Q band. Within the 10 ps-0.45 ns period of time over which our experiments were conducted, the absorbance changes in the Soret and Q bands for MbCO and MbO2 correspond to the conventional long-term, steady-state deoxymyoglobin difference spectra (Mb-MbCO and Mb-MbO2), as determined by comparison of isosbestic, maximum, and minimum points. In addition, MbCO exhibits a decay to a steady state in the Soret band (monitored at 440 nm). The onset of the decay immediately follows photodissociation and has a rate of (8 +/- 3) X 10(9) s-1 (tau = 125 +/- 50 ps). During the 10 ps-0.45 ns observation window, relaxation is not seen for MbO2 in the Soret band, nor is relaxation observed in the Q band for either MbCO or MbO2. We conclude from these results that the steady state that we observed for MbCO and MbO2 is most likely the stable form of deoxymyoglobin, and the relaxational differences between MbCO and MbO2 observed in the Soret band indicate that the electronic destabilization after ligand detachment is very different for these molecules. We believe that these relaxational differences may be related to differences in tertiary structural changes, or due to the fact that the MbCO (S = 0) molecule passes through an intermediate spin Mb (S = 1) state before relaxing the the Mb (S = 2) state.

Animals↗
PubMed1979

Current problems and results in testing microaggregate filters.

Most in vitro studies on microaggregate filters focus on the mechanical retention of microparticles. Data are based on bulk techniques such as differential dry weight and screen filtration pressure measurements or single-particle counting using resistance measurement (Coulter principle). Errors in measurements may result from various steps in sample handling, interaction of the sample with the environment and inborn errors of different techniques. High-speed laser cytometry has been applied and was found to diminish the number of possible errors. The results of these measurements are compared with conventional techniques.

Blood Coagulation↗
PubMed1979

Cell differentiation based on absorption and scattering.

Improvements of the flow system allow calibrated cell length measurements down to less than 2 micron at a very high rate. An optical index match to plane viewing windows perpendicular to the optical axis in the flow system keeps the axial symmetry for forward scattered light. Cell size, axial light loss and scattering intensity within different angles were found to be powerful tools to differentiate cell populations. Red cells were analyzed according to various cell surface structures. Lymphocyte populations isolated from different parts of the lymphatic system in rats have been distinguished. Experimental tumor cells showed typical data pattern after different chemical treatments.

Absorption↗
PubMed1978

Picosecond photodissociation and subsequent recombination processes in carbon monoxide hemoglobin.

Excitation of HbCO by a single 6-psec 530-nm pulse results in photodissociation with a first-order constant of 0.89 X 10(11) sec-1. The kinetics of photodissociation, monitored by following absorbance changes in the Soret band at 440 nm, are interpreted as corresponding to predissociation followed by a corssing into a dissociative state. Subsequent recombination of CO with the porphyrin system and protein structural transformations were monitored by use of a continuous He-Cd laser beam spatially coincident with the photolysis and Soret interrogation beams at the sample. We find that the latter events take place in three distinct time regions, depending on exciation pulse energy and repetition rate. Exictation of HbCO with a single pulse (0.8--5 mJ) results in a relaxation to the ground state with an associative first-order constant of 5 X 10(3) sec-1. With a 100-pulse train (approximately 7.5 mJ), a new decay grows with a rate constant of 63 sec-1. For a pulse-train energy of 12 mJ or higher, a delay occurs at the onset of the second (slower) recombination.

Carbon Monoxide↗
PubMed1976

Fast cell size distribution analysis by laser flow microphotometry-- applications to ciliate populations.

The laser flow microphotometer for rapid cell size analysis is introduced. In contrast to known principles absorption is detected of a part of each cell only. Cell classification is achieved by pulse form analysis of absorption pulses. Cell lengths between 5 and 300 mum have been measured without changing the geometry of the flow system. Other ranges of size can be covered by changing the flow diameters. The principle can be applied to stained or unstained cells. No further preparation of samples is necessary. Examples of high speed measurements on different cell populations of unstained ciliated (Tetrahymena pyriformis), human blood and different pollen grains are given.

Animals↗