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Biomedical subjects

J Vienken

Publications and source records attributed to J Vienken.

71 records · Page 4Linked to original sources
PubMed1984

Magneto-electro-fusion of human erythrocytes.

In inhomogeneous (static) magnetic fields close contact between 'magnetic' human erythrocytes was established. The cells were made magnetic by incubating them in a medium containing small Fe3O4 -particles which adsorbed to the outer membrane surface. Fusion was induced by applying two electric field pulses (field strength: 8.5 kV X cm-1; duration: 60 microseconds) to the magnetically collected cells. This procedure allowed the use of electrically conductive media (3 X 10(-3) omega -1 X cm-1). Fusion of red blood cells occurred very often. If cell suspensions of high density were used fusion resulted in the formation of giant red blood cells with osmotically intact membranes.

Erythrocyte Membrane↗
PubMed1984

Electro-fusion of cells: principles and potential for the future.

Exposure of cells or liposomes to a brief pulse of a strong electrical field can result in a reversible breakdown of the outer membrane. Such breakdown results in an increase in permeability of the plasmalemma, which however re-seals after a short incubation (i.e. the original impermeability is restored). Two or more cells in contact can be made to fuse by this process, provided that the contact is close enough and that the pulse of the electrical field is short enough not to damage the cells. Methods of achieving this contact by electrical and magnetic fields are described. The magnetic method does not demand the use of the low conductivity media used earlier. Other possible modifications of this flexible technique are also described, and used to show how the technique can be modified in future, and how it may be applied to the fields of membrane research, medicine and plant breeding.

Animals↗
PubMed1983

Electrofusion of myeloma cells on the single cell level. Fusion under sterile conditions without proteolytic enzyme treatment.

A technique is presented which allows electrofusion of single cells under sterile conditions. The electrofusion chamber is placed in a Petri dish. Before a droplet of the fusion medium is pipetted between the electrodes, the chamber is completely covered with vaseline, which prevents the fusion medium evaporating. Additionally, the fusion chamber is treated with solutions containing poly(L)-lysine and pronase which results in a decreased movement of the cells on the glass between the electrodes and which allows electrofusion without any proteolytic pretreatment.

Animals↗
PubMed1983

Neutron small angle scattering of matched proteoliposomes with incorporated F0F1 ATPase complex from Rhodospirillum rubrum FR1. An approach to the structure of membrane proteins in their natural environment.

Purified F0F1 ATPase from Rhodospirillum rubrum FR1 has been incorporated into lipid vesicles from the partially deuterated phospholipid dimyristoylglycerophosphocholine (DMPC-D54). These proteoliposomes were able to carry out energy transducing reactions. The incorporation of the membrane protein was controlled by freeze fracture electron microscopy. A method for structural research of the membrane protein in its natural environment has been developed by means of neutron small angle scattering. Using the contrast variation technique, the lipid part of the proteoliposomes was matched by adding an appropriate amount of D2O to the solvent. Thus the neutron scattering profile of F0F1 ATPase incorporated into vesicles was separated from the neutron scattering of the liposome. F0F1 ATPase incorporated in a lipid bilayer, as well as the free enzyme, yields a radius of gyration of Rg = 6.0 +/- 0.1 nm which leads to an overall diameter of 15.5 nm. This result suggests that the monomeric form of F0F1 ATPase is incorporated in DMPC-D54 membranes at 20 degrees C.

Adenosine Triphosphatases↗
PubMed1982

Rotation of cells in an alternating electric field: theory and experimental proof.

Protoplasts of Avena sativa rotate in an alternating electric field provided that at least two cells are located close to each other. An optimum frequency range (20 to 30 kHz) exists where rotation of all cells exposed to the field is observed. Below and above this frequency range, rotation of some cells is only occasionally observed. The angular velocity of rotation depends on the square of the electric field strength. At field strengths above the value leading to electrical breakdown of the cell membrane, rotation is no longer observed due to deterioration of the cells. The absolute value of the angular velocity of rotation at a given field strength depends on the arrangement of the cells in the electric field. A maximum value is obtained if the angle between the field direction and the line connecting the two cells is 45 degrees. With increasing distance between the two cells the rotation speed decreases. Furthermore, if two cells of different radii are positioned close to each other the cell with the smaller radius will rotate with a higher speed than the larger one. Rotation of cells in an alternating electric field is described theoretically by interaction between induced dipoles in adjacent cells. The optimum frequency range for rotation is related to the relaxation of the polarization process in the cell. The quadratic dependence of the angular velocity of rotation on the field strength results from the fact that the torque is the product of the external field and the induced dipole moment which is itself proportional to the external field. The theoretical and experimental results may be relevant for cyclosis (rotational streaming of cytoplasm) in living cells.

Cell Membrane↗
PubMed1980

Erythrocytes and lymphocytes as drug carrier systems: techniques for entrapment of drugs in living cells.

Mouse thymocytes and erythrocytes are loaded electrically with drugs in isotonic solution. The loaded cells are used for targeting the drugs to specific sites in the organism in order to achieve a controlled drug release in time and space. The field technique used for the loading of the cells is based on the dielectric breakdown of the cell membrane which is observed when cell suspensions are subjected to external field pulses of 2-20 kV/cm for short time intervals (ns to microseconds). When an apparent membrane potential of about 1 V is reached in response to the external field, the membrane breaks down reversibly. The breakdown of the membrane is associated with a remarkable and reversible permeability increase of the cell membrane. The increase in permeability depends on the strength and the duration of the field pulse.

Animals↗
PubMed1978

Penetration and entrapment of large particles in erythrocytes by electrical breakdown techniques.

Human erythrocytes suspended in isotonic solutions were subjected to haemolysis by application of an electric field pulse to the cell suspension. The field strengths used were 12 and 16kV/cm, respectively; the pulse duration 40 microseconds. The lysed cells showed resealing properties. The permeability change of the membrane generated by the field pulse and by the subsequent osmotic processes were large enough to facilitate the penetration and entrapment of ferritin and Latex particles (diameter: 0.091 and 0.176 micron, respectively) as revealed by electron microscopy. Correct identification of the Latex particles in the electron-micrographs indicated that LOYTER et al. [J. Cell Biol. 66, 292 (1975)], who recently demonstrated the entrapment of Latex spheres in erythrocytes prepared by osmotic haemolysis mistook electron-dense bodies probably consisting of denaturated protein for Latex particles. Under conditions of osmotic haemolysis, carried out according to BODEMANN and PASSOW, particles could only occasionally be detected within the membrane itself and never within the cell interior, suggesting that the electrical haemolysis method is much more effective in the generation of large holes in the membrane.

Cell Membrane Permeability↗
PubMedDate not supplied

Dialysate contamination and back filtration may limit the use of high-flux dialysis membranes.

Endotoxins, or fragments thereof, can reach the blood stream of dialysis patients, transported by diffusion and connection across the intact high-flux membrane. This transfer depends upon the phenomenon of back filtration. Back filtration generally occurs under conventional high-flux dialysis conditions with membranes having an ultrafiltration coefficient in blood (UF-C) above 20 ml/hr/m2/mmHg. The clinical consequences of back filtration vary from center to center depending primarily on the quality of dialysate. We therefore surveyed the bacterial and endotoxin levels of purified water and effluent dialysate in a cross section of dialysis centers in the central United States. Using a high recovery medium, we found that 53% of the centers had bacterial counts above the Association for the Advancement of Medical Instruments standard in water (20% cfu/ml) and 35% above the standard in dialysate (2,100 cfu/ml). Endotoxin concentrations higher than 5.0 EU/ml in both water and dialysate were found in 4% and 11.8% of the centers, respectively. Since high-flux membranes are believed to be of benefit for long-term dialysis patients, manufacturers will have to offer dialysate preparation systems with additional safety features. The proper membrane design will be a key to the success of such systems.

Colony Count, Microbial↗
PubMedDate not supplied

Rotation of cells in an alternating electric field: the occurrence of a resonance frequency.

Cells suspended in a low-conducting medium were exposed to an alternating electric field whose frequency was altered between 1 kHz and 2 MHz. A resonance frequency was observed at which all suspended cells rotated about an axis normal to the field lines (when the electric field strength was larger than a threshold value of about 400 V/cm). This resonance frequency varied from species to species of cells (mesophyll protoplasts of Avena sativa = 20-40 kHz, human erythrocytes and ghost cells = 80-100 kHz, yeast cells = 140-180 kHz, Friend cells = 30-40 kHz, at room temperature). The resonance frequency of cell rotation was observed only under specific experimental conditions which excluded interference by reversible electrical breakdown of cell membranes and by gravitational forces. Glutardialdehyde fixed and heated cells exhibited no rotation in the frequency and field range investigated. The phenomenon of rotation is discussed in terms of dipole orientation within the membrane.

Animals↗
PubMedDate not supplied

Polymers in dialysis: characteristics and needs.

Polymers employed in dialysis must always be evaluated from the perspective of blood compatibility. This article traces the developments that have taken place in biocompatibility and biostability in dialysis treatment and the mechanisms that are involved.

Biocompatible Materials↗
PubMedDate not supplied

Modified cellulosic dialyzer membranes: an investigative tool in thrombogenicity studies.

We have previously demonstrated that chemical modification of cellulosic membranes with dimethyl-amino-ethyl (DEAE) groups significantly improves membrane properties in terms of biocompatibility. Here, we show that DEAE substitution also alters the membrane's thrombogenic properties, and cellulosic membranes with various amounts of DEAE substitution were produced. Clinical dialyzers were constructed using two experimental membrane materials: modified cellulose-low (MC-low) and MC-high; standard unsubstituted cellulose was used as a control. Six patients were treated for a period of 3 weeks with each type of dialyzer and a heparin dose of less than 6000 IU/treatment. MC-low exhibited less extracorporeal beta-thromboglobulin and thromboxane B2 release than MC-high or Cuprophan. In addition, residual blood volume after clinical use was lower in the MC-low type. MC-low and MC-high induced less complement activation than Cuprophan, as characterized by extracorporeal C5a and C3a plasma concentrations (75% less C5a generation and 50 to 70% less C3a generation than unsubstituted cellulose).

Cellulose↗
PubMedDate not supplied

Successful long-term use of a miniaturized plasmapheresis circuit in rabbits.

Dual lumen silicon rubber right atrial catheters were implanted into the jugular of 8 rabbits and tunneled subcutaneously to exit sites between the ears. Using a miniaturized tubing-pump system, blood flow rates of 25 ml/min could be achieved for up to 3 1/2 hours without sign of hemolysis in an extracorporeal blood circuit. Seven catheters functioned an average of 75 +/- SE 19 days (range 24-117). One catheter remains functional after 176 days. Infection and thrombosis were the main reasons for failure. 20 plasmaphoresis experiments were carried out in four heparinized rabbits (blood flow rates 15 ml/min, plasmaflux 1.5-2.0 ml/min) using polypropylene minifilters (average pore size, 0.55 micron) with the plasma recirculated back into the animal. No hemolysis was detectable throughout the 4 hr experiment. Plasma proteins with a MW of 69 X 10(3) to 3 X 10(6) (Albumin, LDH, SGOT, SGPT, CPK, fibrinogen, LDL) showed a sieving coefficient close to 1.0. The good filtration performance and the absence of side effects make this system a possible use for plasmaphoresis in neonates.

Animals↗
PubMedDate not supplied

Contamination of dialysis water and dialysate. A survey of 30 centers.

The concentration of bacteria and endotoxin in dialysis water and dialysate of 30 dialysis centers in western Germany was examined. Water samples were obtained after treatment by reverse osmosis or other processing methods. Collection of dialysis samples for bacterial, fungal, and endotoxin analysis was conducted before and 2 hours after start of hemodialysis. In 17.8% of all water samples analyzed, the AAMI standard was exceeded and bacterial and fungal counts greater than 200 colony forming units/ml were found. In 11.7% of all dialysate samples, higher contamination than the recommendations for dialysate of 2000 colony forming units/ml were found. The concentration of endotoxin in water and dialysate varied between 0 and 95 endotoxin units in the water samples and 0 and 487 endotoxin units/ml in the dialysate samples. In 12.2% of all water sampled, and 27.5% of all dialysate samples, values of 5 endotoxin units/ml were found. No correlation was found between the level of contamination of either water or dialysate in a specific center and the following factors: water processing method (reverse osmosis or others), type of dialysate (acetate of bicarbonate), type of dialysate machine, or method of machine disinfection. In view of these results it is suggested that endotoxin testing, especially in the dialysate, be a part of regular quality control in dialysis.

Bacteria↗