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Chromaffin granule-microfilament interaction has previously been proposed to play a potential role in the regulation of dopamine uptake into the granules as well as catecholamine secretion in the adrenal medullary cell. However, the microfilament-binding site on the granule surface has not yet been identified. To establish the conditions for solubilization of the binding site, the effects of different-type detergents on the cross-linking of chromaffin granule membranes with F-actin were examined. The cross-linking activity was abolished by treatment of the granule membranes with deoxycholate (DOC), but not Triton X-100 (TX100) and CHAPS. The addition of DOC-extract decreased the viscosity of F-actin solution, suggesting that solubilized proteins bind to F-actin, but not cause the cross-linking of actin filaments. These results indicate that the actin-binding site can be solubilized from chromaffin granule membranes by DOC treatment.

Scanning force microscopy was used for mapping the viscoelastic properties of metaphase chromosomes. These properties were probed by scanning with various imaging forces and subsequent calculation of the difference image. The procedure allows a mapping of the viscoelastic behavior expressed as force-dependent indentation of the local surface feature and results in an image with material contrast. The approach is demonstrated on rehydrated metaphase chromosomes, which were spread and air-dried before rehydration in aqueous buffer. The rehydration resulted in a swelling of the chromosome structure and was accompanied by drastic changes in the viscoelastic properties. For comparisons, force-distance curves on metaphase chromosomes were accumulated; these curves were also used for the calculation of the stiffness curve. The demonstrated approach of mapping viscoelasticity by differential scanning force microscopy allows the detection of domains with varying mechanical properties in biomolecules such as chromosomes.

The effect of bacterial alpha-amylase and an emulsifier on firmness and amylograph characteristics of bread crumb during storage was studied. The bread which contained these additives was less firm and stalled more slowly than the control. The enzyme and the emulsifier retarded firmness but the emulsifier inhibited the action of the alpha-amylase. The regulatory action of the emulsifier also was observed on amylograph characteristics of bread crumb (peak viscosity, cool viscosity and setback).

Treatment with recombinant human deoxyribonuclease I (rhDNase) is currently used as therapy for cystic fibrosis (CF) lung disease. Hypertonic saline (HS) acts as an expectorant promoting mucus secretion and augmenting the volume of sputum. We evaluated the individual and combined effects of HS and rhDNase in vitro on the viscoelasticity of CF sputum. Sputum samples were collected from nine CF patients to use for in vitro testing. Aliquots of CF sputum (0.20 to 0.40 g) were subjected to the following protocols: (1) negative control sample without any treatment; (2) positive control sample, adding 10% volume of normal saline (0.9% NaCl); (3) application of hypertonic saline (HS-3% NaCl); (4) combining approximately 100 nM concentration of rhDNase with protocols 2 and 3. The samples in protocols 2 through 4 were incubated for 30 min at 37 degrees C. For each protocol, CF sputum was analyzed at baseline and at 30 min for spinnability by filancemeter and viscoelasticity by magnetic microrheometry. Spinnability decreased for the sputum samples that were treated with rhDNase, in combination with either HS or normal saline. Treatment with HS alone and combined treatment with rhDNase and HS decreased log G* (the principal viscoelasticity index) to the same degree. Saline alone and rhDNase in normal saline both increased the predicted cough clearability of the sputum; however, the combined treatment with rhDNase and hypertonic saline had the best overall effect on cough clearability. The change in predicted mucociliary clearability, although greatest after HS, was not significant. These in vitro results suggest that combined treatment with rhDNase and HS should be evaluated further as a potential mucotropic approach to augment the clearance of purulent sputum in CF lung disease.

Erythromycin inhibits mucus glycoconjugate secretion from airway cells in vitro and may increase mucus clearance in patients with asthma or diffuse panbronchiolitis. Because mucus hypersecretion is common in purulent rhinitis, we questioned whether clarithromycin would change the properties of nasal mucus from subjects without sinus disease and from patients with acute purulent rhinitis. Nasal secretions were collected before and after nasal methacholine challenge from 10 adults without nasal symptoms and without methacholine from 10 patients with purulent rhinitis. After 2 wk of oral clarithromycin (500 mg twice daily), secretions were again collected from both groups. Secretions were analyzed for viscoelasticity, cohesion, hydration, and ciliary and airflow (sneeze) transportability. Compared with secretions from healthy subjects, rhinitis secretions had decreased wettability (contact angle on Teflon 100 degrees versus 84.67 degrees; p = 0.001), increased cohesion (36.8 versus 24.3 mm; p = 0.003), decreased sneeze clearability (20.6 versus 32 mm; p = 0.04), and increased percent solids (4.61 versus 2.82%; p = 0.04). After clarithromycin, the rheology, hydration, cohesion, and transportability of the rhinitis secretions were similar to those of the postclarithromycin secretions from the healthy subjects. Secretion volume also decreased (500.1 versus 28.3 mg; p = 0.01), and mucociliary transportability increased by 30% (0.76 versus 0.99; p = 0.005). Although clarithromycin reduced mucus secretion in both rhinitis patients and normal subjects, it did not alter the secretory response to methacholine.

Using micropipette aspiration technique, the authors investigated the viscoelastic properties of human fetal hepatocytes (HFH) and hepatocellular carcinoma (HCC) cells. And further, the effects of two cytoskeleton interferents, cytochalasin D and colchicine on the viscoelastic properties of HFHs were investigated. The results showed: the elastic coefficients K1 and K2 of HCC cells were significantly higher than those of HFHs; the action of cytochalasin D led to an obvious decrease in elastic coefficients K1, K2 as well as the viscous coefficient mu of HFHs; under the action of colchicine, the K1 value of HFHs increased while the K2 and mu values decreased obviously or tended to decrease. A brief discussion was presented about these results.

The inflammatory reaction in the human gastric mucosa to Helicobacter pylori could be initially triggered by an array of cytokines expressed in infected gastric epithelial cells. The spiral morphology and flagella of these organisms could increase their velocity in a viscous environment such as methylcellulose solution. The goal of this study was to determine whether modification of H. pylori motility could influence the expression of cytokine genes from gastric epithelial cells infected with H. pylori. Adherent human gastric epithelial cells were cultured and overlaid with methylcellulose solutions of varying viscosity. These epithelial cell layers covered with methylcellulose solution were inoculated with H. pylori. RNAs were then extracted from the gastric epithelial cells. Various cytokine gene expressions were assessed and quantified by reverse transcription-polymerase chain reaction (RT-PCR) and standard synthetic RNA. Cytokine proteins were also measured by enzyme-linked immunosorbent assay (ELISA). Expression of mRNA for interleukin(IL)-8 was upregulated in H. pylori-infected gastric epithelial cells overlaid with methylcellulose of 15 centipoise (cp) viscosity. The expression of mRNA for IL-1 alpha, IL-8, monocyte chemotactic protein (MCP)-1 and granulocyte macrophage colony-stimulating factor (GM-CSF) was also upregulated in H. pylori-infected gastric epithelial cells overlaid with methylcellulose solution of the same viscosity. The number of molecules of the expressed cytokine transcripts also paralleled the amounts of protein secreted from gastric epithelial cells infected with H. pylori. These results suggest that methylcellulose solution (simulating the mucus layer in vivo) could increase contact of H. pylori with gastric epithelial cells by increasing its motility. This could result in the upregulation of mRNA for proinflammatory cytokines in gastric epithelial cells, therefore enhancing inflammatory reaction at H. pylori-infected sites.

The decrease in membrane microviscosity of erythrocyte ghosts in the presence of clinically relevant concentrations of seven inhalation anaesthetic agents was studied using fluorescence polarization anisotropy of the membrane incorporated fluorescent probes 1,6-diphenyl-1,3,5-hexatriene and 1-[4-trimethylammoniumphenyl]-6-phenyl-1,3,5-hexatriene. All anaesthetic agents produced a dose-dependent decrease in anisotropy of both probes, indicating decreased membrane microviscosity. The reduction in anisotropy measured at the minimum alveolar concentration (ED50) for anaesthesia was related inversely to the anaesthetic potency of the agent and was directly proportional to the hypothetical concentration of agent in the membrane calculated from lipid-water partition coefficients. These findings do not support the hypothesis that volatile anaesthetic agents act by increasing membrane microviscosity of the bulk lipid bilayer to produce anaesthesia.

Cystic fibrosis (CF) patients develop progressive cytokine-mediated inflammatory lung disease, with abundant production of thick, tenacious, protease- and oxidant-rich purulent airway secretions that are difficult to clear even with physiotherapy. In the search for a potential treatment, we have tested tyloxapol, an alkylaryl polyether alcohol polymer detergent previously used as a mucolytic agent in adult chronic bronchitis. Tyloxapol inhibits activation of the transcription factor nuclear factor-kappa B (NK-kappa B), reduces resting secretion of the cytokine interleukin-8 (IL-8) in cultured human monocytes, and inhibits lipopolysaccharide (LPS)-stimulated release of tumor necrosis factor-alpha (TNF-alpha), IL-1 beta, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), and the eiconsanoids thromboxane A2 and leukotriene B4 (LTB4). We have previously shown that tyloxapol is a potent antioxidant for hydroxyl radicals ( OH). Tyloxapol (0.05 to 0.1% wt/vol) effectively scavenges the oxidant hypochlorous acid (HOCl; 1 to 7.5 mM) in vitro, and protects from HOCl-mediated lung injury in rats. Tyloxapol also reduces the viscosity of CF sputum (from 463 +/- 133 to 128 +/- 52 centipoise). We conclude that tyloxapol is potentially useful as a new antiinflammatory therapy for CF lung disease, and could possibly promote clearance of secretions in the CF airway.

Background: The use of acetylcysteine as a mucolytic agent is controversial. In 1962 it was claimed to be "the most effective agent ... for the liquefaction of ... secretions" but was subsequently taken off the market as a respirator solution. Normal saline, on the other hand, is becoming increasingly popular. A study was undertaken to determine which solution is more effective at clearing retained secretions following thoracotomy., Methods: The study included 10 patients and was of single blind, two-way crossover design. Measurements taken before and after each treatment included sputum viscosity, difficulty of expectoration, weight of sputum expectorated, and oxygen saturation., Results: Following nebulisation of acetylcysteine, sputum viscosity was reduced, difficulty of expectoration was reduced, the weight of sputum expectorated was increased, and oxygen saturation was increased. There were no changes after nebulisation of normal saline., Conclusions: This study shows that, following thoracotomy, nebulised acetylcysteine reduces sputum viscosity, making expectoration easier and improving oxygenation. Nebulised normal saline has no effect.

Background: In vitro studies have shown that pH is an important determinant of mucus structure and function, but the relationship in vivo is unclear. Omeprazole increases intragastric pH and also allows bacterial overgrowth. In this study we have assessed the effect of omeprazole on gastric juice viscosity and examined the influence of pH and bacterial overgrowth on the resulting change., Methods: Gastric juice specific viscosity, pH and total bacterial counts were measured in nine healthy male volunteers before and after omeprazole 20 mg twice daily for 1 week. The effect of incubation at pH 2 and 7 was also determined. Viscosity changes were compared with changes in pH and bacterial counts., Results: Mean viscosity fell (P < 0.05) following treatment, though there was a wide range in viscosity both before and after treatment. The decrease was reproduced by incubation of pre-treatment juice at pH 7 but not pH 2. The fall in viscosity was correlated (P < 0.05) with change pH., Conclusion: Omeprazole decreases gastric juice, and hence gastric mucus, viscosity by increasing intragastric pH. This could be important if it allows improved penetration of antimicrobials to Helicobacter pylori within the mucus layer.

1. Old rats showed a significant decrease in the number of muscarinic M(1) receptors and a significant increase in membrane microviscosity in the striatum and hippocampus as compared to young animals. In contrast, no significant changes in the density of muscarinic M(2) receptors were observed with aging. 2. Chronic treatment of aged rats with L-alpha-glycerylphosphorylcholine (L-alpha-GPC) restored the number of M(1) receptors to levels found in the striatum and hippocampus from young animals. The same treatment to aged rats partially restored membrane microviscosity in both regions studied and hence increased membrane fluidity. 3. None of the major metabolites of L-alpha-GPC (choline, glycerophosphate or phosphorylcholine) was able to restore the number of striatal and hippocampal M(1) sites and membrane microviscosity of aged rats, neither did any of these treatments (including treatment with L-alpha-GPC) modify the level of M(1) receptors and microviscosity values in young rats.

A newly synthesized mucolytic agent, N-acetylcysteine L-lysinate (Nacystelyn) was studied. Tracheal mucus velocity (TMV), transepithelial potential difference (PD), rheological properties, and ion content of collected airway secretions were evaluated in six healthy mongrel dogs after placebo, Nacystelyn (NAL) and acetylcysteine (NAC) metered dose inhaler (MDI) aerosols. Although TMV was increased and viscoelasticity decreased after both treatments, the treatment effect with NAL was significantly greater. Furthermore, NAL increased the negative PD and CI- content of secretions in the trachea, an effect not observed after NAC. Both compounds increased ciliary beat frequency (CBF) on the frog palate at a concentration range similar to that approximated in dog airways. The increased mucociliary clearance could be partially explained by favourable rheological changes combined with stimulation of CBF. Since both compounds break disulfide bonds in mucus polymers, the greater change in mucus rheology and clearance rate after NAL, without change in water content, could be explained by the increase in CI- content. Nacystelyn appears to combine different modes of action which synergistically cause an increase in the clearance rate of airway secretions.

Albumin reduces capillary hydraulic conductance (Lp) even at low concentrations. To determine if part of this barrier protective effect might be extracellular, we studied the effects of bovine serum albumin (BSA) on Lp of self-assembled basement membrane (Matrigel). Lp with tris(hydroxymethyl)aminomethane (Tris) buffer superfusate was stable at 1.77 +/- 0.22 x 10(-5) (SE) cm.s-1.cmH2O-1 over several hours. At 0.1 g/dl BSA, experimental/control (Tris) Lp fell to 83.1 +/- 6.0% (2P < 0.025), with decreases to 72.4 +/- 3.7% at 1 g/dl (2P < 0.005), 45.3 +/- 5.1% at 2.5 g/dl (2P < 0.001), and 45.0 +/- 4.8% at 4.0 g/dl (2P < 0.001). In separate experiments, BSA arginine groups were neutralized by 1,2-cyclohexanedione (CHD), and experimental/control Lp values were measured. At 2.5 g/dl, CHD-BSA depressed Lp to 54.4 +/- 4.8%, while unmodified BSA reduced Lp to 40.8 +/- 3.5% of Tris control (2P = 0.05). Finally, soluble arginine at three- and sixfold the arginine in BSA was added to BSA superfusate. For threefold, Lp rose to 120 +/- 8% of BSA level and for sixfold to 129 +/- 9% (2P < 0.05). We conclude that some part of the albumin protective effect is very likely due to consequences on extracellular matrix and that at least 18-22% of this effect is related to arginine groups on albumin when computed from Lp, and up to 34% when viscosity is taken into account. Membrane-saturable arginine-binding sites can be unbound with arginine, thus nullifying part of the barrier protective effect of BSA.

The changes in the structure and properties of erythrocyte membranes that are induced by free fatty acids and their derivatives have been studied. The state of the membrane has been evaluated using the activity of membrane-bound acetylcholinesterase (AChE), the pyrene monomer/excimer fluorescence intensity ratio as an indicator of membrane lipid microviscosity and the fluorescence of membrane-bound 1-anilinonaphtalene-8-sulphonic acid (ANS). Free fatty acids and corresponding aliphatic aldehydes induced an inhibition of membrane-bound AChE, effectively decreased the bulk lipid and protein-bound lipid microviscosity, and quenched the fluorescence of membrane-bound ANS. The type and efficiency of the enzyme inhibition, as well as the efficiency of microviscosity decrease and ANS fluorescence quenching, depended on the hydrophobicity and the end group in the effector molecule. Therefore, it is proposed that fatty acids and related compounds perturb the lipid bilayer and disturb the protein-lipid complementarity of the human erythrocyte membrane.

A new one-step computational procedure is presented for estimating the parameters of the nonlinear three-element windkessel model of the arterial system incorporating a pressure-dependent compliance. The data required are pulsatile aortic pressure and flow. The basic assumptions are a steady-state periodic regime and a purely elastic compliant element. By stating two conditions, zero mean flow and zero mean power in the compliant element, peripheral and characteristic resistances are determined through simple closed form formulas as functions of mean values of the square of aortic pressure, the square of aortic flow, and the product of aortic pressure with aortic flow. The pressure across as well as the flow through the compliant element can be then obtained so allowing the calculation of volume variation and compliance as functions of pressure. The feasibility of this method is studied by applying it to both simulated and experimental data relative to different circulatory conditions and comparing the results with those obtained by an iterative parameter optimization algorithm and with the actual values when available. The conclusion is that the proposed method appears to be effective in identifying the three-element windkessel even in the case of nonlinear compliance.

EPR spin-probe method was used to investigate the dependence of erythrocyte cytosol microviscosity (ECM) on the concentration of different substances and temperature. The critical ECM value has been determined after reaching of which erythrocytes become destroyed. It is shown that this ECM value is reached at different NaCl concentrations depending on the temperature.

Normal tendons, normal palmar aponeuroses and specimens from patients with Dupuytren's disease, namely apparently normal palmar aponeuroses and contracture bands were subjected to elastase and chondroitinase ABC digestion. Maximum Young's modulus, normalized hysteresis loop and residual elongation were determined before and after enzyme treatment. In normal tendons, normal and apparently normal palmar aponeuroses both normalized hysteresis loop and residual elongation increased significantly after elastase treatment, whereas the stiffness decreased. Normalized hysteresis loop and residual elongation display changes corresponding to the amount of digested elastin. The increased viscosity of untreated contracture bands containing less elastin, as compared to normal palmar aponeurosis, was not affected by elastase. Obviously, the elastic fibers in normal shape and distribution are responsible for maintaining an elastic status with a low viscous stress component. With the exception of contracture bands chondroitinase ABC caused a minor increase of residual elongation and as opposed to elastase a decrease of normalized hysteresis loop indicating an increased mobility of the tissue fibers.

Purpose: To evaluate circulatory alterations induced by intra-arterial injection of iodized oil and emulsions of iodized oil with an anticancer drug., Materials and Methods: The viscosities and stability were evaluated in arterial flow of iodized oil and the emulsions. Doppler ultrasound was used to quantify the embolic effect in the arteries of rabbits. The behavior of doxorubicin and iodized oil in the rat cremaster muscle was studied with videomicroscopy., Results: The emulsions did not break up after injection, and the size of discontinuous-phase droplets did not change. The embolic effect did not correlate with viscosity. The thinnest water-in-oil emulsion had the lowest embolic effect and induced homogeneous distribution of iodized oil droplets in the arterial tree., Conclusion: Changes in the formulation of emulsions obtained with the same proportions of drug and iodized oil change the distribution of iodized oil in the arterial tree, the location at which the drug is released, and the embolic effect.

1. The mechanism of interaction of the polyacrylates, carbopols with the mucus barrier in vivo has been investigated in vitro. 2. Carbopol caused a dramatic increase in the viscosity of porcine gastric mucin solutions that was up to 19-fold greater than that of the sum of the individual polymers. 3. The mucin-carbopol interaction was stable after an initial 30 min period for up to 36 h at 25 degrees C or 37 degrees C. It was reduced by increasing the temperature from 20 degrees C to 45 degrees C, was unaffected by pH and ionic strength, but was enhanced by Ca2+. 4. The magnitude of the interaction between mucin and carbopol depended on the polymeric structure of the mucin and the molecular size and level of cross-linking of the carbopol. 5. The interactions were reversible and increased with increasing carbopol and mucin concentration. The dramatic increase in viscosity can be explained in terms of space filling by the mucin molecules leading to predominantly carbopol-carbopol interactions. 6. Carbopol 934P inhibits pepsin hydrolysis and therefore has potential as a mucosal protective agent in vivo.

MR889 is a synthetic inhibitor of human neutrophil elastase with potential in clinical conditions characterized by a high load of this proteolytic enzyme, mainly chronic obstructive pulmonary disease. On the basis of its mechanism of inhibition, it has been suggested that MR889, upon reaction with elastase, would generate new free thiol groups. The aim of this study was to investigate whether MR889, upon reaction with elastase, may modify rheologic properties, i.e. apparent viscosity and elasticity, of both human sputum and porcine stomach mucus, in ex-vivo and in vitro experiments. MR889 10(-4) M alone had no effect on rheologic properties of samples, whereas an agent with free thiol group, n-acetylcysteine 10(-4) M reduced both viscosity and elasticity. MR889 10(-4) M upon reaction with elastase 0.03 nMol (giving 83% of elastase inhibition) had no effect on rheologic properties of samples, whereas upon reaction with elastase 0.15 nMol (49% inhibition) yielded a marked decrease in viscosity. This result would suggest that MR889, in the presence of a high load of elastase, may acquire direct mucus modifying capacity, likely because of its greater utilization upon reaction with the enzyme.

The influence of ozone and sunlight in a concerted reaction on hyaluronate solutions was investigated. The kinematic viscosity of hyaluronate solutions is decreased by ozone-air mixtures and simultaneous radiation with sun rays within a few minutes, indicating a depolymerization of the hyaluronate molecule. The reaction is dependent on the concentration of ozone and on the time of exposure to ozone and sunlight. The concerted degradation of hyaluronate is more effective than the reaction with each component, ozone and sun rays, alone. We conclude that hyaluronate depolymerization by ozone and sunlight may be one factor for irritations of the eye by photochemical smog and increased exposure to sun rays.

1. The effects of an indigestible soluble polysaccharide (carboxy methyl cellulose: CMC) on broiler performance (body weight gain, food and water intake) and on chyme characteristics (moisture content, viscosity, pH, osmolality and retention time) in broilers were studied. 2. In semi-synthetic diets 0, 5.0 and 10.0 g/kg of cellulose was replaced by CMC on weight basis. These diets were fed to male broilers from 3 to 5 weeks of age. 3. When 10.0 g/kg CMC was included in the diet, food intake and body weight gain were reduced, compared to the 0, and 5.0 g/kg CMC diets. Food:gain ratio and water intake were increased at each CMC concentration. The significant quadratic response showed an increased response per 5.0 g/kg dietary CMC at the higher CMC concentration. 4. The viscosity in the supernatant of the chyme was linearly increased in all intestinal segments by CMC. A quadratic increase was observed in the crop. In the lower ileum, differences between the 5.0 and 10.0 g/kg CMC diets were not significant. 5. CMC increased the mean retention time of chromium in the duodenum and in the upper jejunum, and reduced the maximal rate of marker excretion. The transit time (first appearance of the marker in the excreta), however, was significantly reduced at the highest CMC concentration. 6. The osmolality of the intestinal fluid decreased less as the chyme moved from the duodenum into the lower ileum, with increasing concentrations of CMC. 7. The ileal pH was reduced linearly by the CMC content of the diet. 8. Based on a higher moisture content of the chyme in the CMC-fed birds, and the higher jejunal and ileal osmolarities in those birds, it was concluded that the efficiency of both digestion and absorption was reduced by CMC inclusion in broiler diets.

Viscosupplementation is a new medical concept that has as its therapeutic goal the restoration of rheological homeostasis in pathological structures such as osteoarthritic joints. When the normal viscoelasticity of a solid tissue compartment or the elastoviscosity of a liquid tissue compartment is decreased under pathological conditions, normal function and regenerative processes are impaired. By introducing viscosupplementary devices, the normal rheological state of such compartments is restored or augmented. These devices stay in the tissue compartment for various periods of time, depending on the nature of the viscosupplement and the pathophysiology of the tissue compartment.

Intestinal ischemia is considered a major factor in the development of necrotizing enterocolitis (NEC). Despite this, the majority of affected infants lack documentation of clinical events associated with obvious gut hypoperfusion. Recent evidence in adults suggests that endotoxin may impair flow in the microcirculation through alterations in erythrocyte deformability. As the gut serves as a semipermeable reservoir of endotoxin in the stressed neonate, such localized activity may result in intestinal ischemia at the microcirculatory level through alterations in the red cell membrane. This study evaluates the role of endotoxin on neonatal erythrocyte membrane viscosity. Paired anticoagulated whole blood specimens were obtained from the umbilical cord of 10 neonates at delivery. Samples were incubated with either 2 micrograms/mL of E coli endotoxin (LPS) or an equal volume of saline (control). Following incubation, erythrocytes were isolated, washed, and incorporated with the fluorescent membrane probe TMA-DPH. Membrane viscosity was assessed by spectroscopic analysis of the fluorescent emissions induced by excitation of the probe at 365 nm. Results were calculated as anisotropy and analyzed for differences by ANOVA. Endotoxin resulted in a significant increase in red cell membrane viscosity as compared to control (LPS 291.2 +/- 5.1 v Control 271.7 +/- 3.3, P < .01). As the effects of endotoxin are known to be primarily the result of white blood cell (WBC) activation, this study was repeated in an additional 10 neonates in whom WBCs were removed prior to endotoxin/saline incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Solutions of purified brevin were applied to skinned thin bundles or isolated fibres of smooth muscle. This produced a sharp drop of isometric tension, an effect due to the severing effect of brevin on actin filaments, partially depleted from tropomyosin in skinned preparations. On skinned single fibres, brevin accelerates the speed of unloaded shortening. As no effect was detected on the myofibrillar ATPase turnover rate, brevin was thought to affect the viscosity of the cytoplasm. This was confirmed by analysis of the cytoplasm stiffness which decreased in the presence of brevin. It is proposed that Ca-activated brevin acts on actin-filamin gels, set in parallel to the contractile apparatus.

The relative viscosity of Pseudomonas aeruginosa alginate was shown to increase markedly when combined with mucin, Ca2+ ions and the exopolysaccharide from Pseudomonas cepacia. The presence of such a heterodisperse polysaccharide solution significantly reduced the diffusion and hence antimicrobial activity of tobramycin and to a lesser extent ciprofloxacin against Ps. aeruginosa by factors of 90 and 2.5-fold respectively over a 5 h incubation period. The clinical implications of these results are discussed in relation to cystic fibrosis.

Inositol phospholipid turnover is enhanced during mitogenic stimulation of cells by growth factors and the breakdown of phosphatidylinositol 4,5-bisphosphate (PtdInsP2) may be important in triggering cell proliferation. PtdInsP2 also binds actin-binding proteins to regulate their activity, but it is not yet understood how this control is achieved. The protein alpha-actinin from striated muscle contains large amounts of endogenous PtdInsP2, whereas that from smooth muscle has only a little but will bind exogenously added PtdInsP2. In vitro alpha-actinin binds to F-actin and will crosslink actin filaments, increasing the viscosity of F-actin solutions. We report here that alpha-actinin from striated muscle is an endogenous PtdInsP2-bound protein and that the specific interaction between alpha-actinin and PtdInsP2 regulates the F-actin-gelating activity of alpha-actinin. Although the F-actin-gelating activity of alpha-actinin from smooth muscle is much reduced compared with that from striated muscle, exogenous PtdInsP2 can enhance the activity of smooth muscle alpha-actinin to the level seen in striated muscles. These results show that PtdInsP2 is present in striated muscle alpha-actinin and that it is necessary for alpha-actinin to realize its maximum gelating activity.

Nasal decongestants are widely used in the treatment of rhinitis. Decongestant drugs are one of the main components, but numerous reports gave been made concerning side, local and general effects. The problem is particularly important in the treatment of infantile rhinitis for which the use of saline solution alone is often preferred. The paper evaluates the fluidifying action of a saline solution and polysorbate 20, a tensioactive molecule, on nasal mucous. Changes in mucous viscosity were assessed using a "filancemeter" and thromboelastograph. At the same time a clinical study on the efficacy of treatment and the possible presence of side effects was carried out in a population of rhinitic subjects. The study highlighted the fluidifying effect of the preparation on nasal mucous and the complete absence of side effects.

Paraquat (PQ) is a widely used herbicide that causes acute adult respiratory distress syndrome (ARDS) and chronic lung damage (diffuse fibrosis). One of the earliest biochemical effects induced by PQ is damage to type II pneumocytes with consequent depletion of surfactant. With the aim of counteracting the toxic effects of PQ, a series of investigations were performed into the possible protective effect of the drug ambroxol, which induces the synthesis of surfactant in lung alveolar type II cells. The number of survivors and survival time of rats treated ip with 35 mg PQ/kg was significantly increased by 3 days of ambroxol pretreatment and by ambroxol treatment 30 min or 2 hr after PQ. Total phospholipid content in lung and bronchoalveolar lavage fluid (BALF) was significantly reduced 30 hr after treatment with PQ alone. The association of ambroxol with PQ significantly antagonized this reduction. In BALF the ratio between palmitic acid and stearic acid concentrations was significantly lower in animals treated with PQ alone but was returned to normal by the association with ambroxol. The cell line A549, exposed in vitro to PQ concentrations from 0.5 x 10(-4) to 2 x 10(-3) M, showed a significant dose-dependent loss of viability. Cells pretreated with ambroxol (10 mg/ml) were more resistant to PQ and their viability started to decrease significantly only from a PQ concentration of 0.8 x 10(-3) M. Membrane microviscosity was measured on the same cells. Cells treated with PQ alone showed a reduction of membrane microviscosity, which was significantly counteracted by ambroxol pretreatment. The curves of modification of membrane microviscosity of cells treated with PQ and with ambroxol plus PQ paralleled those of cell viability, indicating that the stimulation of surfactant synthesis in vitro may be a prerequisite for counteracting some of the early effects of PQ.

1. The gastroprotective properties of a new H2-receptor antagonist, ebrotidine, against ethanol-induced mucosal injury was investigated. 2. Groups of rats, with and without indomethacin pretreatment, received intragastrically either a dose of ebrotidine or vehicle only, followed by ethanol given at various intervals up to 4 hr. The gastric mucosa, 30 min after the ethanol challenge, was then subjected to macroscopic and histologic examination, and physicochemical measurements. 3. Ebrotidine at doses of 50 mg and higher per kg body wt effectively prevented the alcohol-induced mucosal injury, even in the presence of indomethacin. The protective effect was demonstrable already at 30 min, reached maximum at 1 hr, and persisted up to 3 hr. 4. Physicochemical analyses established that ebrotidine elicited 30% increase in mucus gel dimension, caused 19-20% increase in glycolipids and phospholipids, and evoked 21% increase in sulfomucin and 18% in sialomucins. As a consequence, the mucus gel viscosity increased by 1.4-fold, H+ retardation capacity by 16%, and hydrophobicity by 65%. 5. The results demonstrate that ebrotidine is a unique H2-antagonist endowed with a remarkable mucosal strengthening capability.

We studied the protective effect of prostaglandin E1 (PGE1)-induced hypotensive anesthesia on gastric mucosa in 30 elective surgical patients. Three groups, each composed of 10 patients, received PGE1, nitroglycerin or none during general anesthesia. Then we measured the viscosity and pH of gastric fluid continuously in each group. In the PGE1 group the viscosity and pH increased significantly and rapidly (P less than 0.05) as compared with the other groups. This suggests that PGE1 offers prophylactic effect against postoperative acute gastric mucosal lesion (AGML).

The influence of an oversulphated glycosaminoglycan, pentosanpolysulphate, on hyaluronan metabolism of the synovial lining cell was studied in vivo in human volunteers. Significant increases in the mean degree of polymerisation of the hyaluronan chains were observed after a series of four to six intra-articular injections of this glycosaminoglycan. No increases in hyaluronan synthesis rates were observed. Repeated administration of the drug did not cause any inflammation or bleeding in the joint cavity.

Blood samples from ten patients with clinically evident manifestations of atherosclerotic disease were used to evaluate in vitro the effects of exogenous glutathione (GSH) on platelet aggregation and on blood filtration and viscosity. In two groups, each of ten atherosclerotic patients, matched for sex, age, prevalent localization, duration and gravity of disease, the effects of GSH infusion (600 mg every day for seven days) on the haemocoagulative pattern, platelet aggregation and blood filtration and viscosity were evaluated in a double-blind, placebo cross-over trial. The GSH in vitro addition resulted in a significant increase in blood filtration and a significant decrease in blood viscosity and platelet aggregation. The venous GSH infusion both significantly decreased blood viscosity and increased blood filtration. Partial thromboplastin time was lengthened after GSH infusion even thought it remained in the normal range.

Since one of the cellular targets of cetiedil, a vaso-erythroactive drug, is likely to be the erythrocyte membrane, we have studied the influence of this drug on erythrocyte membrane microviscosity and acetylcholinesterase activity. No effect was evidenced on microviscosity, as measured by fluorescence polarization of light emitted by DPH or TMA-DPH labelling of the lipid bilayer. Cetiedil, however, did lower acetylcholinesterase activity, but it did not directly inhibit this enzyme activity. It can therefore be considered as an amphiphilic drug that perturbs membrane properties without affecting the physical state of the erythrocyte membrane.

Using a viscometric assay and various electron microscopic procedures (negative staining, rotary shadowing, ultrathin sectioning) we have determined the influences of different kinds of ions and of ionic strength on the structures formed by assembly of soluble subunits of vimentin from bovine lens tissue or from Escherichia coli transformed with Xenopus vimentin cDNA. In contrast to the assembly of typical, i.e., 8 to 14-nm, intermediate-sized filaments (IFs) at elevated (e.g., 160 mM) concentrations of monovalent cations and at millimolar Mg2+ concentrations, filaments formed in the presence of Ca2+ ions (e.g., 5 mM) appeared at a lower rate, attained lower viscosity and were considerably thicker and shorter. The largest diameter measured was that for the recombinant amphibian protein: 24.2 +/- 8.5 nm in negative staining, 28.7 +/- 5.6 nm in sections. These thick Ca(2+)-induced filaments, however, revealed the same approximately 2 nm protofilament composition and approximately 20 nm cross-striation pattern as typical IFs, indicative of a similar molecular arrangement. The significance of this unusual structural IF protein assembly is discussed.

Membrane microviscosity, phospholipid composition, and turnover were measured in cultured vascular smooth muscle cells isolated from mesenteric arteries of stroke-prone spontaneously hypertensive and age-matched, normotensive Wistar-Kyoto rats. Membrane microviscosity, measured with fluorescence polarization, revealed greater microviscosity (lower fluidity) of the membranes isolated from smooth muscle cells from hypertensive as compared with those isolated from normotensive rats (p less than 0.01). Preincubation of membranes from hypertensive rats with 5 mM calcium reduced membrane microviscosity in "core" and in "surface" regions of the bilayer toward values observed in Wistar-Kyoto rats. Phospholipid composition did not differ between intact aortas and cultured mesenteric cells or between those tissues obtained from normotensive and from hypertensive rats. The total lipid-associated radioactivity was significantly lower in cells from stroke-prone spontaneously hypertensive rats than in those from Wistar-Kyoto controls (p less than 0.01). Phosphatidylcholine incorporated 70% and phosphatidylinositol 16% of total lipid-associated radioactivity, with no difference between cells from hypertensive and normotensive animals. Turnover of phosphatidylethanolamine was greater in cells from Wistar-Kyoto rats (p = 0.02), whereas turnover of phosphatidylserine was greater in cells from stroke-prone spontaneously hypertensive rats (p = 0.04). The greater microviscosity of the lipid bilayer in hypertension is a generalized defect of the matrix in which the transport proteins function. We hypothesize that this defect is responsible for the multiple abnormalities of membrane transport systems that have been described in genetic hypertension.

Effect of Erythropoietin (Ep) on the interaction of Concanavalin A (Con A) with rat erythrocytes was studied using 125I-labelled Con A. Binding of Con A to erythrocytes was dependent on time and cell concentration. Starvation caused an elevation of the lectin binding capacity of red cells which again came down towards the normal level on Ep administration to starved rats. Binding of Con A to erythrocytes decreased linearly with increasing concentration of Ep. Specificity of binding was confirmed by inhibition studies with alpha-methyl-D-mannopyranoside (Me Man) Cells from the starved rats compared to those from normal and Ep treated animals were less prone to inhibition by this sugar analog. Positive cooperative binding of Con A to rat erythrocyte was observed at low concentration of Con A but was absent at higher lectin concentrations. Starvation caused an increase in the number of binding sites per cell which returned to normal level after Ep treatment. Under identical conditions, binding affinities were not much changed in these cells. Cells from the starved animals were more susceptible to agglutination compared to those from normal and Ep-treated rats. Microviscosity and cholesterol/phospholipid ratio of red cell membrane decreased in the starved animals which retraced its way back towards the normal level after Ep treatment.

Developing a method for isolating skate (Raja erinacea) basolateral liver plasma membranes, as well as characterizing the lipid composition and fluidity of these membranes, was the primary purpose of this study. Membranes were isolated using self-generating Percoll gradients. Marker enzyme studies indicate that this preparation is highly enriched in the basolateral domain of the liver plasma membrane and largely free of contamination by intracellular organelles or canalicular membranes. Further, these membranes contain the agency responsible for Na(+)-dependent alanine transport. This finding indicates that this membrane preparation will be useful for the study of skate liver plasma membrane transport processes. The lipid composition and fluidity (as assessed by the fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene) of the skate basolateral liver plasma membrane shows little variation among preparations. Further, DPH anisotropy plotted as a function of temperature yields a straight line (r = 0.99) which indicates that there is no lipid phase change in these membranes from 4 degrees to 37 degrees C. The membrane preparation does contain substantial phospholipase A2 activity. The function of this enzyme is, in part, to modify membrane lipid composition and fluidity in response to temperature variations; therefore, this finding suggests that in situ lipid metabolizing enzymes may play a central role in the adaptation of skate basolateral liver plasma membranes to changes in the ambient temperature.

The effects of hydroxypropylmethylcellulose on upper gastrointestinal transit, viscosity, and water flux were studied in six dogs fistulated at the proximal duodenum and/or mid-jejunum. Combinations of different grades of hydroxypropylmethylcellulose were prepared as 2% or 3.3% solutions to yield input viscosities of low (approximately 5000 cp at 37 degrees C and 1 s-1), medium (15,000 cp), or high (30,000 cp) viscosity. Hydroxypropylmethylcellulose modified intralumenal viscosity, with a linear relationship existing between input and lumenal viscosity. With regard to transit, the lag time before the onset of chyme recovery increased linearly as a function of luminal viscosity. There was also a pronounced decrease in the first-order emptying rate constant as lumenal viscosity increased from water to low-viscosity hydroxypropylmethylcellulose, but as viscosity was further increased there was little additional change. These results indicate that water-soluble fibers can exert a significant influence on both the lumenal viscosity and the transit profile in the upper gastrointestinal tract.

The effect of cyclosporine A (CsA) on peripheral blood lymphocyte (PBL) membranes was studied using fluorescence techniques. Light scattering and intercalation experiments indicate CsA has a critical micelle concentration of 3 x 10(-5) M. Doses above this critical concentration were avoided during these in vitro studies. Steady-state fluorescence polarization of the lipid probe DPH in peripheral blood lymphocyte membranes revealed that 10(-5) M CsA caused a significant increase in the fluorescence polarization 1-5 min after exposure (15-16% above control DPH polarization values). This initial increase in polarization plateaued at 10-11% above control values within roughly 40 min. Fluorescence lifetime measurements of DPH during this response reveal a slight increase in DPH lifetime. The increased DPH lifetimes showed that polarization measurements alone underestimate the rigidity of DPH's microenvironment. Measurements of DPH polarization and lifetime were analyzed using the Perrin equation to calculate the apparent microviscosity. Control PBL membranes at 37 degrees C exhibited a microviscosity (eta) equal to 1.89 poise (P). Five to 10 min after CsA exposure eta = 2.62-2.68 P and plateaus at eta = 2.31-2.51 P 20-120 min after exposure. Dose-response studies show that prolonged (greater than 90 min) exposure to CsA alters lymphocyte membrane fluidity. This early response in lymphocytes to CsA may correspond to other events associated with the initial binding of CsA to lymphocyte membranes such as membrane depolarization and alterations in phospholipid metabolism. Intravenous delivery of CsA in clinical studies has shown that a similar CsA dose of near 10(-5) M is available in the blood during the first hour after injection. The evidence presented here suggests that peripheral blood lymphocyte membranes are rigidified when exposed to CsA doses similar to those found in patient's blood immediately following intravenous administration.

A strain of Agrobacterium sp. S-1231 was isolated from one of the soil samples taken from Beijing area. In 4 days' cultivation it was found to produce extracellular polysaccharide and about 24 grams per liter of sugar-containing substrate. At the same concentrations, the polymer produced higher viscosity than that of xanthan gum (Kelzan). The viscosities of 0.2%, 0.5% and 1.0% solutions of this polymer were found to be 840, 5200 and 15000 cp respectively, measured with a Brookfield LVF viscosimeter at 6 rpm. Besides its pseudoplastic rheological properties and the excellent stability towards pH, it had unusual compatibility with high levels of salts and provided shear thinning properties similar to that in fresh water. Moreover, it is also compatible with cationic dyes without precipitation. Unlike other microbial polysaccharides, the viscosities of this polymer remained constant at different temperatures under 65 degrees C, but it decreased sharply in a narrow range of temperatures between 70-75 degrees C.

The apparent viscosity of three test meals containing 6% of different guar gums was measured in the gastric, duodenal and jejunal contents and related with the gastrointestinal myoelectric activity and transit time in pigs. The motility index (mV/min) and the spike burst frequency (number of spike bursts/min) were closely related to the digesta viscosity: they were significantly increased by most viscous contents in stomach and small intestine. This effect was associated with an increase in the orocecal transit time of liquids and solids.