Physiological NaCl Solution Research Articles

Gonadotropin-releasing hormone treatment improves locomotor activity, urinary function and neurofilament protein expression after spinal cord injury in ovariectomized rats

It was reported that the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH) produces neurotrophic effects and that the spinal cord possesses GnRH receptors. The aim of the present study was to determine whether administration of GnRH improves locomotor activity, urinary function and neurofilament (NFs) protein expression after spinal cord injury (SCI) in ovariectomized rats. SCI was induced by balloon inflation model resulting in paraplegia. Locomotion was evaluated according to the Basso, Beattie, and Bresnahan Scale. Rats were subjected to bladder compression, twice daily until bladder reflex was established. NFs of 68, 160 and 200kDa from spinal cords were analyzed by electrophoresis. GnRH (60μg/kg) or physiologic NaCl solution was administered at 1 day after SCI and then daily for 15 days and the functional evaluation was realized for 5 weeks. Our results indicate that locomotor activity, restoration of urinary dysfunction and NFs expression of 160 and 200kDa were improved in SCI animals given GnRH compared to those without treatment. These findings suggest that GnRH acts as a neurotrophic factor and may be used as a potential therapeutic agent for treatment of SCI.

Longitudinal analysis of taurine induced effects on the tear proteome of contact lens wearers and dry eye patients using a RP-RP-Capillary-HPLC–MALDI TOF/TOF MS approach

Tear proteomic studies revealed distinct similarities between contact lens wearers and dry eye patients. AMO Complete® multipurpose contact lens cleaning solutions containing taurine seem to have a beneficial effect regarding contact lens induced dry eye. To illuminate the effect of taurine on the tear proteome of contact lens wearers and sicca patients we developed a gel-based RP-RP capillary HPLC-MALDI TOF/TOF MS strategy. Two contact lens wearer groups, one using eye drops containing 0.05% taurine; the other for control physiological NaCl solution were monitored. Also, a third group of sicca patients using taurine solution was studied (N=4 individuals/group). Tear pools of each group at six time points over 5 weeks were analyzed. In summary 267 tear proteins were identified. We found a protein subset showing a linear taurine response with R(2) values ≥0.5. Taurine effects were detected predominantly in the contact lens group demonstrated by distinct level decreases. Most protein candidates were related to inflammation. Since levels of these proteins differentiate from those of a healthy non-contact lens wearer reference they are supposed to be involved in contact lens induced dry eye and should be focused on in further studies.

The protein binding substance ibuprofen does not affect the T1 time or partition coefficient in contrast-enhanced cardiovascular magnetic resonance.

BackgroundContrast enhanced cardiovascular magnetic resonance (CMR) with T1 mapping enables quantification of diffuse myocardial fibrosis. Various factors, however, can interfere with T1 measurements. The purpose of the current study was to assess the effect of co-medication with a typical protein binding drug (Ibuprofen) on T1 values in vitro and in vivo.Methods50 vials were prepared with different concentrations of gadobenate dimeglumine, Ibuprofen and human serum albumin in physiologic NaCl solution and imaged at 1.5T with a spin echo sequence at multiple TRs to measure T1 values and calculate relaxivities. 10 volunteers (5 men; 31±6.3 years) were imaged at 1.5T. T1 values for myocardium and blood pool were determined for various time points after administration of 0.15mmol/kg gadobenate dimeglumine using a modified look-locker inversion-recovery sequence before and after administration of Ibuprofen over 24 hours. The partition coefficient was calculated as ΔR1myocardium/ΔR1blood, where R1=1/T1.ResultsIn vitro no significant correlation was found between relaxivity and Ibuprofen concentration, neither in absence (r=−0.15, p=0.40) nor in presence of albumin (r=−0.32, p=0.30). In vivo there was no significant difference in post contrast T1 times of myocardium and blood, respectively and also in the partition coefficient between exam 1 and 2 (p>0.05). There was good agreement of the T1 times of myocardium and blood and the partition coefficient, respectively between exam 1 and 2.ConclusionsContrast enhanced T1 mapping is unaffected by co-medication with the protein binding substance Ibuprofen and has an excellent reproducibility.

The effect of silver chloride formation on the kinetics of silver dissolution in chloride solution

The precipitation and growth of AgCl on silver in physiological NaCl solution were investigated. AgCl was found to form at bottom of scratches on the surface which may be the less effective sites for diffusion or the favorable sites for heterogeneous nucleation. Patches of silver chloride expanded laterally on the substrate until a continuous film formed. The ionic transport path through this newly formed continuous film was via spaces between AgCl patches. As the film grew, the spaces between AgCl patches closed and ion transport was primarily via micro-channels running through AgCl patches. The decrease of AgCl layer conductivity during film growth were attributed to the clogging of micro-channels or decrease in charge carrier concentration inside the micro-channels. Under thin AgCl layer, i.e. on the order of a micrometer, the dissolution of silver substrate was under mixed activation-Ohmic control. Under thick AgCl layer, i.e. on the order of tens of micrometers, the dissolution of silver substrate was mediated by the Ohmic resistance of AgCl layer.

Treatment of Clinically Diagnosed Equine Sarcoid with a Mistletoe Extract (Viscum album austriacus)

Equine sarcoids (ES) are common, difficult to treat, and have high recurrence rates. Viscum album extracts (VAE) are used in human cancer treatment. That therapy with VAE (Iscador P) is effective in the treatment of ES. Fifty-three horses (444 ES); 42 were treated with VAE or placebo as monotherapy; 11 were treated with VAE or placebo after selective excision of ES. Prospective, randomised, blinded, clinical trial. Horses were randomly assigned to treatment (VAE; n=32) or control group (Placebo; n=21). One milliliter of VAE (Iscador P) in increasing concentrations from 0.1 to 20 mg/mL or physiological NaCl solution was given SC 3 times a week over 105 days. Number, localization, and type of the ES were documented over 12 months. A subset of 163 clinically diagnosed equine sarcoid (CDES) lesions (95 VAE, 68 Placebo) was evaluated in detail, considering clinical findings and tumor volume. No undesired adverse effects were observed except for mild edema at the injection site in 5 of 32 horses (16%). Complete or partial regression was observed in 13 horses of the VAE group (41%) and in 3 of the control horses (14%; P<.05). After VAE treatment, 48 of 95 CDES (67%) showed an improvement compared with 17 of 68 CDES in the control group (40%; P<.01). Twenty-seven CDES had disappeared completely in the VAE group (38%) compared with 9 CDES in the control group (13% NS). VAE (Iscador P) represents a safe and effective treatment for CDES.

Treatment of equine sarcoid with the mistletoe extract ISCADOR® P (viscum album austriacus) – a double-blind placebo controlled study

Equine Sarcoid (ES) is the most common skin tumour in horses with high recurrence rate. No single universal treatment is effective. Viscum album extracts (VAE) are used as adjunct in the treatment of human cancer. We hypothesized that therapy with VAE (Iscador® P) is also effective in Equine Sarcoids. Of 53 horses with ES, 42 were treated solely with VAE or placebo as monotherapy and 11 were treated after selective excision of ES. Horses were randomly assigned to the VAE (n=32) or placebo group (n=21). Horses received increasing concentrations of VAE from 0.1mg/ml to 20mg/ml or physiological NaCl solution 3 times a week over 105 days s.c. Number, localization, size and type of the ES were documented over 12 months. Cure or improvement rate (decrease of tumour size by 50% or more in at least half of the tumours) were assessed after one year. Sarcoid level assessment included analysis of 1 to 7 clinically relevant sarcoids per horse. In the VAE group, 13 horses (41%) showed improvement. Of these, 9 patients showed complete remission (28%). In the control group only 3 cases (14%), all showing complete remission, were classified as improved. In the VAE group 27 ES (38%) showed complete remission and 48 (67%) improvement after one year compared to 9 ES with complete remission (13%; n.s.) and 17 ES (40%; p&lt;0.01) with improvement in the placebo group. VAE (Iscador® P) represent a safe and effective treatment for ES, particularly in cases with multiple sarcoids.

A New Method of Synthesis of Aspartic Acid/Lactic Acid Copolymers Under Microwave Field

Aspartic acid/lactic acid copolymers were synthesized under microwave radiation, without a catalyst, by bulk and solution method. The reaction was carried out also in the presence of phosphoric acid as the catalyst and with the use of a conventional heating. Furthermore, partial degradation of the copolymers by hydrolysis in a physiological NaCl solution and in the Lactated Ringer's solution was investigated. The structure of the products was characterized by Fourier-transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1H NMR). Thermalproperties of the aspartic acid/lactic acid copolymers were determined by thermogravimetry (TG) and differential scanning calorimetry (DSC), while their molecular weight was measured by gel permeation chromatography (GPC).

Mechanical properties of a degradable phosphate glass fibre reinforced polymer composite for internal fracture fixation

This paper reports on the mechanical properties and pH upon degradation of phosphate glass fibre reinforced methacrylate-modified oligolactide. Phosphate glass fibres of the composition 51.04 P 2O 5–21.42 CaO–25.51 Na 2O–2.03 SiO 2 (mol%) were produced by a crucible spinning technique. Fibres were embedded into a matrix of a degradable organic polymer network based on methacrylate-modified oligolactide; samples with and without addition of CaCO 3 for pH control were produced. pH during degradation in physiological NaCl solution could be increased to up to 6.5 by addition of 20 wt.% calcium carbonate to the fibre composites. pH in Tris buffer solution was about 7.11. Mechanical properties of dry specimens were investigated during 3-point bending tests and gave elastic moduli in the range of cortical bone (15 to 20 GPa). However, addition of calcium carbonate decreased tensile strength of the fibre composites and resulted in brittle fracture behaviour, while CaCO 3-free composites showed a fibrous fracture mode. Control of pH and degradation is a requirement for obtaining a fracture fixation device with degradation properties matching in vivo requirements. Results show that addition of CaCO 3 is suitable for controlling the pH during degradation of metaphosphate glass polymer composites.

Getaktete Dreifachapplikation mit perfundierten Nadelapplikatoren bei Radiofrequenzablation: In-vitro-Versuch an der Rinderleber / Pulsed radiofrequency ablation using perfused needle applicators in an in vitro trial on bovine liver

Intermittent energy application seems to have positive effects in achieving necrotic zones. We analyzed different pulse periods (PPs) to optimize this method. A radiofrequency alternating current was delivered via a triple-needle applicator (3 cm distance of each needle) introduced into freshly procured bovine liver. The open applicator system was constantly perfused with physiological NaCl solution (3×80 ml/h, power output was constant 90 W). Radiofrequency current was fed to the individual needle in turn of varying PPs (1, 2, 5, and 7 s) over 15 min. Each experimental run comprised a total of 30 applications and temperature was recorded over time. The largest necrotic diameters were found at PP 1 s (relevant: shortest axial diameter/D in the center of the lesion: 9.27 cm, SD±0.97 cm) (PP 2 s D=8.65 cm, SD±0.95 cm, p=0.021; PP 5 s D=8.35 cm, SD±0.95 cm, p=0.001; PP 7 s D=8.18 cm, SD±0.79 cm, p=0.0001). Neither doubling the perfusion rate nor raising the concentration of the perfusion liquid led at PP 1 s to increased necrotic diameters (p=0.82). Our study shows that reducing the PP to 1 s of an open perfused intermittent radiofrequency ablation system produces significantly larger coagulation volumes in explanted liver tissue reaching necrotic diameters over 9 cm. Neither doubling perfusion rates nor higher concentrated perfusion liquid increase necrotic diameters.

Interaction of DNA/Polycation Complexes with Phospholipids: Stabilizing Strategy for Gene Delivery

The interaction between negatively charged lipid vesicles and positively charged DNA/polylysine complexes was studied. The interaction does not lead to release of DNA from the initial complexes. The particles formed are easy to prepare, they have slight negative charge, small dimensions and show good stability in physiological NaCl solution. Such properties might indicate that stabilization of the particles by lipid coating might be a potent strategy, alternative to PEGylation of DNA/polycation complexes. Interaction of DNA/polycation complex particles with lipid vesicles.

Effect of thyroliberin on the course of experimental candidosis in mice

Thyroliberin (TRH) is one of the hormones, which affect immunologic processes. This hormone was studied in experimental subacute candidosis in mice. BALB/c males were given intraperitoneally single dose of Candida albicans cells (1 x 10(7) to 1 x 10(9) cfu of strain no. 244-33 ATCC). The animals from the experimental groups were injected subcutaneously, after 24 h from inoculation, dose of 10 microg TRH in 0.2 ml 0.9% NaCl, seven times at 24 h intervals. The control animals were given respectively 0.2 ml of physiologic NaCl solution. We have found that the examined hormone significantly decreases mortality in these animals (LD50 C. albicans for mice treated with TRH was three times higher than that in the control groups), prolongs mean survival time for mice and decreases the intensity of fungal invasion of the animal organs.

Interactions of different types of localized corrosion in surgical implants.

Surgical implants often show different types of localized corrosion such as corrosion fatigue cracking, pitting and crevice corrosion on the same part. Interactions of these different corrosion phenomena were investigated. This was done by cyclic loading of electropolished tensile specimens at different constant and changing potentials. Material investigated was a surgical implant steel X2CrNiMo18-15-3 which was immersed in physiological NaCl solution. Pitting and repassivation potentials were determined. Samples with and without artificial cracks as well as masked specimens were tested. Incubation period for first damage, density and size of pits by coulometric and volumetric method were determined. The fracture surfaces were then investigated by SEM. Results show that not in all cases pitting corrosion was the cause for corrosion fatigue cracking. Also pitting is favoured by crack formation. Density of pits increases by a factor of 5 without any change to pitting potential. There are primary pits formed prior to crack initiation and secondary pits formed after crack initiation. At samples without crack there is almost no difference between the optically measured value of total pit volume and the coulometrically determined value. At samples with cracks coulometric volume of pits is much larger than optical one. This proves that there is a significant amount of crevice corrosion in the crack. The corrosion current density in the crack increases by two orders of magnitude when comparing it to electropolished surface of the sample. Results of laboratory experiments are confirmed by failure of a real implant.

Influence of shot peening on the corrosion fatigue properties of a stainless steel for surgical implants

AbstractFor some applications in surgical implants a rough surface is desired, since the jagged surface structure assists the adhesion of bone material or surface coatings. Some implants are peened with coarse and sharp edged peening media. In order to avoid a distinct loss of fatigue strength, spherical peening balls should be used. The present paper shows for stainless steel X2CrNiMo18‐15‐3 that the effect of surface roughening can be compensated by the effect of compressive residual stresses in the surface layer.However, the effect of surface flaws must not be ignored. For this reason, the shape and depth of surface flaws in shot peened surfaces has been examined and a classification of different types of surface flaws is presented.Fatigue tests in laboratory air and in physiological NaCl solution showed that there is a distinct influence of the depth and shape of surface flaws on the fatigue strength. The use of coarser peening balls and a subsequent electrolytic polishing treatment leads to an increase of fatigue strength.Furthermore, it is shown that shot peening may have a positive or negative effect on corrosion fatigue strength.

Die Kastration männlicher Lämmer mittels Immunisierung gegen GnRH

The objective of this study was to evaluate the effect of a GnRH-vaccine in the ram lamb. Experiments were performed using 20 male lambs, randomly divided into a test (GnRH-immunization) and control group (physiological NaCl-solution). At a body weight of 20 kg (age 2-3 months) and three weeks later, all animals of the test group received 2 ml of Improvac (CSL Limited, Parkville, Victoria, Australia). The body weight as well as the blood testosterone concentration were measured weekly for 16 weeks. Thereafter, blood samples for testosterone analysis were taken monthly in immunized lams only. After the booster injection testicular growth was suppressed and plasma testosterone remained at low values < 0.1 ng/ml for at least 12 weeks. The mean corresponding testosterone concentrations for the control lambs ranged between 0.1 and 0.9 ng/ml plasma. An increase of testosterone was observed in 8 of 10 immunized animals between 3 to 7 months after the booster dose. The control lambs showed a tendency for better growth rate than vaccinated animals, but the difference was not significant. Our results demonstrate that in prepubertal ram lambs two immunizations with Improvac, three weeks apart, can suppress testosterone secretion and testicular growth at least for three months after the booster injection. For a suppression of reproductive function longer than three months after the second vaccination, a third immunization is needed at this time or when testicular growth is beginning.

A new method to optimize the adhesion between bone cement and acetabular bone in total hip arthroplasty

In cemented total hip arthroplasty cup loosening occurs earlier than stem loosening in most of the cases. This is mainly caused by the lack of stabile adhesion between the hydrophobic bone cement and the hydrophilic bone surface of the acetabulum. The aim of this study was to develop a multilayer amphiphilic bonding system which prevents the hydrolytic debonding at the bone cement-bone interface, thus optimizing the compound stability. In a first series of tests a standardized three-point-bending test was performed to determine the compound stability of the bone cement-bone interface. The bony test specimens were immersed into physiologic NaCl solution to simulate the hydrolytic in situ conditions and contaminated with blood to simulate the intraoperative bleeding of prepared acetabular bone surfaces. In a second series of tests polyethylene cups were implanted into the acetabular cavity of sheep using the current cementing technique. Acetabular bone stock was prepared differently (subchondral sclerotic zone preserved vs. removed, additional drilling into the acetabular roof, with vs. without the multilayer bonding system) in intra-individual comparison of both acetabular sides. To ascertain the bone cement-bone stability a torsional-turn out test was performed on an universal testing machine. In the three-point-bending tests the compound stability between bone cement and bone was 50- to 100-times higher with the use of the multilayer system. In the torsional-turn out tests the compound stability showed in mean a 1.8-fold increase of the interface strength in case of preconditioned acetabular cavities with the multilayer bonding system. The developed multilayer bonding system optimizes the interface strength between acetabular bone stock and bone cement significantly for cemented cups in total hip arthroplasty. In contrast to cementing techniques with complete removal of the subchondral sclerotic zone (in order to optimize micro-interlocking) the biologically effective and load bearing acetabular bone stock can be preserved using the newly developed multilayer bonding system. This aspect might be highly important especially with regard to possible acetabular bone defects caused by the process of aseptic cup loosening.

FACTORS INFLUENCING THE EFFICIENCY OF LIPOPLEXES MEDIATED GENE TRANSFER IN LUNG AFTER INTRAVENOUS ADMINISTRATION*

The objectives of this study were to test the influence of different parameters on the in vivo cationic lipid mediated gene transfer in lung after intravenous administration. Luciferase activity was evaluated in lung tissue 24 hours after intravenous administration of different types of lipoplexes. These included lipoplexes prepared using cationic phosphonolipids or DOTAP and various amounts of plasmid DNA. Using two different plasmids we tested the influence of plasmid size on transfection efficiency in vivo. In a last series of experiments, lipoplexes were prepared using different excipients (water, NaCl or 5% glucose solution) and three injection volumes were tested. We demonstrate that chemical structure modifications such as cation substitution and increment of the aliphatic chain length significantly improve transfection efficiency. High luciferase levels are obtained by increasing lipid to DNA charge ratio and plasmid DNA dose and decreasing plasmid size. Lipoplexes prepared in physiological NaCl solution and injected using a volume of 800μl are significantly the most effective.Cationic lipid mediated gene transfer in lung tissue after intravenous administration is influenced by factors including cationic lipid chemical structure, lipid to DNA ratio and plasmid dose. Nevertheless, plasmid size, injection volume and the excipient, used for the lipoplexes preparation, are also important factors and must be considered for an optimization of in vivo gene delivery using intravenous administration.

Low power integrated pressure sensor system for medical applications

A new low power integrated pressure sensor system with digital output (1 bit PDM signal) for medical applications is presented. The absolute pressure sensor comprising 400 nm thick surface micromachined polysilicon membranes for capacitive pressure detection and a monolithic integrated 2nd order sigma–delta-modulator including voltage reference and timing generator is extremely miniaturized on an area of approximately 3 mm 2. For protection and biocompatibility reasons the sensor is coated with a silicone elastomer of up to 100 μm thickness, which does not influence the sensor's performance. The sensor system was tested in vitro in physiological NaCl solution, showing excellent results compared to a commercial available reference sensor. The sensor system is working well down to a supply voltage of 2.2 V with a power consumption of 0.5 mW. The resolution is better than 12 bit. Due to the small chip area, low power consumption and cost effective production process, the sensor is ideal for medical applications, e.g., in combination with telemetric power and data transmission [J. Zacheja, B. Clasbrummel, J. Binder, U. Steinau, Implantable Telemetric Endosystem for Minimal Invasive Pressure Measurements, MedTech95, Berlin, Germany, (1995)] as an implantable sensor to reduce the mortality risk of intensive care patients.

Renal phosphate excretion in goats.

It was the aim of these experiments to characterize further the role of salivary glands and kidneys in phosphate (Pi) homeostasis of small ruminants. Renal clearance experiments were performed in five female, non-pregnant adult goats during infusions of physiological NaCl or isotonic phosphate solution. Basal plasma Pi concentrations of 1.6 +/- 0.3 mM (NaCl infusion) were associated with a renal resorption of the filtered Pi by 98.5-99.3% and Pi was excreted in the urine at a rate of 1.2-2.8 mumol/min. Elevating the plasma Pi levels up to 7.2 +/- 1.7 mM resulted in significant increases of Pi excretion. The mean plasma threshold for renal Pi excretion of these animals was high (4.3 +/- 1.0 mM) and thus substantially higher than the physiological range of plasma Pi. The maximal tubular resorption rate for Pi in goats was found to be higher than the respective values in monogastric animals. In relation to the enhanced Pi concentrations in plasma the salivary Pi concentrations increased simultaneously and reached a maximum level of about 66.3 mM at plasma levels between 4-6 mM. Indications for hormonal regulation mechanisms via parathyroid hormone (PTH) and calcitriol could not be detected during the infusion periods.

Stabilization of small unilamellar DMPC-liposomes by uncharged polymers

Polymer-free and polymer-bearing small unilamellar (SUV) liposomes from dimyristoyl-phosphatidylcholine (DMPC) were prepared under standardized conditions. Polymer-bearing liposomes were formed by incorporating an uncharged polymer [hydrolyzed poly(vinyl alcohol) (PVA), poly(vinyl alcohol-co-vinylacetal) (PVA-Al), poly(vinyl alcohol-co-vinyl propional) (PVA-Prol) poly(vinyl alcohol-co-vinyl butiral) (PVA-Bul) copolymer or poly(vinyl pyrrolidone) (PVP)] into the membrane bilayer of vesicles. The kinetic (long-term) stability of the liposome dispersions stored in distilled water, in physiological NaCl solution and at various pH values, respectively, were studied. The physical stability of vesicles was tested by measuring the size and the zeta potential of liposomes by means of a Malvern Zetasizer 4 apparatus. It was shown that most of these polymers are effective steric stabilizers for the DMPC-liposomes. Among the polymers, the PVA-Bul and PVA-Prol copolymers and the PVP of high molecular mass exhibited the most efficient stabilizing effect at each pH studied, indicating that the formation of a relatively thick polymer layer around the lipid bilayers ensures an enhanced and prolonged physical stability of liposomes. Also, the butiral or propional side chain in the PVA-based copolymers presumably promotes the anchoring of macromolecules to the vesicles. Using these macromolecules, the colloidal interactions between vesicles can be modified and so the physical stability of liposomes and the kinetic stability of liposome dispersions can also be controlled.

Bending undulations and elasticity of the erythrocyte membrane: effects of cell shape and membrane organization

The undulatory excitations (flickering) of human and camel erythrocytes were evaluated by employing the previously used flicker spectroscopy and by local measurements of the autocorrelation function K (t) of the cell thickness fluctuations using a dynamic image processing technique. By fitting theoretical and experimental flicker spectra relative values of the bending elastic modulus Kc of the membrane and of the cytoplasmic viscosity eta were obtained. The effects of shape changes were monitored by simultaneous measurement of the average light intensity I0 passing the cells and by phase contrast microscopic observation of the cells. Evaluation of the cellular excitations in terms of the quasi-spherical model yielded values of Kc/R3(0) and mu.R0 (R0 = equivalent sphere radius) and allowed us to account (1) for volume changes, (2) for effects of surface tension and spontaneous curvature and (3) for the non-exponential decay of K (t). From the long time decay of K (t) we obtained an upper limit of the bending elastic modulus of normal cells of Kc = 2-3 x 10(-19) Nm which is an order of magnitude larger than the value found by reflection interference contrast microscopy (RICT, Kc = 3.4 x 10(-20) Nm, Zilker et al. 1987) but considerably lower than expected for a bilayer containing 50% cholesterol (Kc = 5 x 10(-19) Nm, Duwe et al. 1989). The major part of the paper deals with long time measurements (order of hours) of variations of the apparent Kc and eta values of single cells (and their reversibility) caused (1) by osmotic volume changes, (2) by discocyte-stomatocyte transitions induced by albumin and triflouperazine, (3) by discocyte-echinocyte transitions induced by expansion of the lipid/protein bilayer (by incubation with lipid vesicles) and by ATP-depletion in physiological NaCl solution, (4), by coupling or decoupling of bilayer and cytoskeleton using wheat germ agglutinin or erythrocytes with elliptocytosis and (5) by cross-linking the cytoskeleton using diamide. These experiments showed: (1) Kc and eta are minimal at physiological osmolarity and temperature and well controlled over a large range of these parameters. (2) Echinocyte formation does not markedly alter the apparent membrane bending stiffness. (3) During swelling the cell may undergo a transient discocyte-stomatocyte transition. (4) Strong increases of the apparent Kc and eta after cup-formation or strong swelling and deflation are due to the effect of shear elasticity and surface tension. Our major conclusions are: (1) The erythrocyte membrane exhibits a shear free deformation regime which requires ATP for its maintenance.(ABSTRACT TRUNCATED AT 400 WORDS)