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Flow dichroism, flow polarized fluorescence and viscosity of the DNA acridine complexes.

The strong binding of various acridine dyes to DNA has been studied by the measurements of flow dichroism, flow polarized fluorescence and viscosity. Negative flow dichroism and percentage change in polarized fluorescence intensity show that intercalated dye molecules are oriented rather perpendicularly to the main axis of the DNA helix, like base pairs. On the other hand, viscosity measurements show that the increase of the contour length of DNA depends on the dye structure, being much smaller in the case of dyes with bulky substituents compared to that of the other dyes. This may be attributed to the formation of the outside bound complex. Further, the introduction of bulk substituents to the acridine ring leads to a little smaller values of the reduced dichroism and intensity change of polarized fluorescence. The results may be qualitatively understood if we assume that the outside bound dye lies in the groove of the DNA helix and the plane of the dye tilts from the perpendicular direction relative to the main axis of the helix.

Acridines↗

A computerized system for acquiring DNA solution viscosity data.

A new computerized system for monitoring time-dependent viscosity changes of DNA solutions is reported. It provides an extremely sensitive measure of DNA lesions induced by chemical carcinogens. The described apparatus consists of some precision mechanical modules, derived by an early manual version of the system, and of some new electronic parts as optical sensors, a microprocessor-based unit and a personal computer. The new system allows the analysis of more than one specimen at the same time. Compared with the early manual method of measuring, this computerized system, by removing subjectivity and a certain number of casual and systematic errors that might occur with the operator manual intervention, makes possible the evaluation of the resolving power of the viscometers themselves in detecting viscosity changes.

DNA↗

The use of amniotic fluid viscosity measurements to establish fetal lung maturity.

A Haake viscosimeter was used to determine the viscosity of amniotic fluid and this value correlated with fetal lung maturity. The results were evaluated along the weeks of pregnancy and compared to those obtained using Gluck's method (lecithin/sphingomyelin ratio (L/S). Using 91 separate amniotic fluid samples, free of meconium and blood contamination, we found that the new method was quick and easy to perform. Thirty-one neonates were delivered within 48 h of the amniotic fluid sampling and the percentage of correct prediction of respiratory distress syndrome (RDS) was 71% with a mature L/S ratio and 83% with viscosity less than 1.17 cP. In cases with an immature L/S ratio (value below 2) the correct prediction of RDS was 64%; with the fluorescence polarization (FP) technique it was 91%. We conclude that the amniotic fluid FP value is a reliable index of fetal lung maturity and risk for developing RDS and has also specific technical and diagnostic advantages over the L/S ratio.

Amniotic Fluid↗

Plasma viscosity and biochemical parameters in the "fatty" rat.

The essential part played by rheological factors in the genesis of thrombosis and atherosclerosis has often been mentioned. Thus, we undertook an investigation on plasmatic rheological factors in an animal model with modifications in plasma lipids (homozygotic and heterozygotic "fatty" rats) compared to normal animals (Sprague-Dawley rats). Test parameters were: cholesterol, triglycerides, lipidograms, glucose, total proteins, fibrinogen, plasmatic viscosity. Experiments gave the following results in homozygotic animals as compared to heterozygotic rats and controls: hyperlipidemia with massive increase in triglycerides, hyperfibrinogenemia, increase in total proteins and glucose, modifications in lipidogram in the form of more lipoproteins migrating in the beta-position and fewer proteins migrating in the alpha-position. Such changes were accompanied by an increase in whole blood and plasma viscosity. After considering the various correlations between these parameters, the authors suggest the application of such an animal model to atherogenesis.

Animals↗

Effect of oral administration of highly purified eicosapentaenoic acid on platelet function, blood viscosity and red cell deformability in healthy human subjects.

Eicosapentaenoic acid (EPA), which is abundant in seafood, has been reported to be a potent antagonist of platelet aggregation and also to reduce the incidene of cardiovascular disorders. We recently reported that EPA also reduces whole blood viscosity. A highly purified EPA, in a soft capsule (75% ethylester form of EPA; EPA-E), manufactured from sardine oil was administered to 8 healthy male subjects for 4 weeks. No side effects were observed. Platelet aggregation and platelet retention significantly decreased. The EPA content in platelet phospholipids markedly increased but docosahexaenoic acid (DHA) and arachidonic acid (AA) contents did not change. A reduction in whole blood viscosity and an increase in erythrocyte deformability were also observed after 4 week's ingestion of EPA-E. The EPA content in erythrocyte membrane phospholipids markedly increased after 4 weeks, and was positively correlated with erythrocyte deformability. Reduction of platelet aggregation and improvement of the rheological properties of the erythrocyte might be explained by an increase in the EPA content in platelet and erythrocyte phospholipids.

Administration, Oral↗

Direct influence of blood viscosity on pulmonary vascular resistance.

When cardiac output is maintained constant in the intact animal, changes in blood viscosity exert a direct and rapid effect on pulmonary vascular resistance. The presence of intact cardiopulmonary reflex mechanisms does not alter the direct influence of blood viscosity on pulmonary vascular resistance.

Animals↗

The influence of perfusate viscosity, RBC deformability and drag on the function of an isolated perfused rat kidney.

There is increasing clinical interest in improving blood rheology to optimize organ function, but studies correlating the two are scarce. To study this, rat kidneys were perfused in vitro at 37 degrees C at a constant mean renal arterial pressure of 160 mm Hg. The perfusate consisted of an oxygenated Krebs HCO3 buffer containing 1 mg/ml glucose, 0.5 mg/ml creatinine, amino acids, [3H]inulin (marker for GFR), 2.5 g/dl albumin, and 10 or 20% hematocrit. In some experiments, RBC were made nondeformable by heating at 50 degrees C for 20 min. Deformability was measured by an ektacytometer. In other experiments, 0.001% of an anionic polyacrylamide (Separan), a drag-reducing agent, was added to the perfusate. Viscosity was measured with a cone and plate viscometer, [Na+] with a flame spectrophotometer, and perfusate flow with a Brooks in-line flowmeter. Other functional parameters, GFR, urine flow, RPF, and reabsorption of sodium and water, were also measured. The results indicate that (1) making RBC nondeformable or increasing viscosity by increasing hematocrit reduces renal function, and (2) addition of a drag-reducing agent improves renal function at 20% hematocrit. We conclude that rheologic manipulation of a perfusate solution can alter flow and renal function.

Absorption↗

Is maternal whole blood viscosity a factor in fetal growth?

Many authors have correlated changes in maternal hemodynamics during pregnancy with fetal growth. Sufficient plasma volume expansion in pregnancy seems mandatory for an optimal reproductive result. A correlation between rheological characteristics (low-shear whole blood viscosity and yield shear stress) and birthweight centiles was found in 26 third-trimester pregnancies. The level of statistical significance (P less than 0.05) was reached in nulliparous pregnancies but not in parous pregnancies. Measurements of whole blood viscosity at low shear rate and yield shear stress seem to provide information on the efficacy of placental perfusion. There have been reports in the literature to support the assumption of the prevalence of low-shear circumstances in the intervillous space. The hypothesis is put forward that, during pregnancy, the changes in maternal hemodynamics influence fetal growth by their impact on the flow through the placenta.

Adult↗

Effects of the non-selective beta-adrenoceptor blocking agent, carteolol, on platelet function, blood coagulation and viscosity.

We have studied the effect of a new beta-adrenergic blocker, carteolol, on platelet function, blood coagulation and viscosity in 10 healthy male volunteers. Following carteolol (5 mg orally) we were able to demonstrate significant inhibition of platelet aggregation to ADP (p less than 0.05) and adrenaline (p less than 0.01) after 5 hours, but not at 2 or 24 hours. This maximum inhibition of platelet aggregation corresponded to peak plasma concentrations of carteolol measured. The platelet release reaction, as measured by plasma levels of the platelet specific protein beta-thromboglobulin (BTG) was unaltered and there was no significant effect on a panel of coagulation tests or on blood viscosity.

Adenosine Diphosphate↗

Correlation of the internal microviscosity of human erythrocytes to the cell volume and the viscosity of hemoglobin solutions.

The microviscosity of the cytoplasm of human erythrocytes as well as of membrane-free hemoglobin solutions was investigated measuring the rotation of the small spin-label molecule, Tempone. The dependence of the intracellular microviscosity on the extracellular pH and osmotic pressure which was varied by NaCl or sucrose was sufficiently explained on the basis of alterations of the red blood cell volume. The intracellular microviscosity depended exclusively on the hemoglobin concentration. It did not differ from that of comparable membrane-free hemoglobin solutions. It was not necessary to take into account long-range interactions between hemoglobin molecules. The conclusion therefore was that the intracellular viscosity is not modified by cytoplasmic structures or the cell membrane. Above a hemoglobin concentration of 6 mM the viscosity of hemoglobin solutions increased much faster than the microviscosity. From measurements obtained with different spin-labels it followed that also the charge of these molecules is of importance.

Cyclic N-Oxides↗

High viscosity hydroxypropylmethylcellulose reduces postprandial blood glucose concentrations in NIDDM patients.

The ability of high viscosity hydroxypropylmethylcellulose (HPMC) to reduce postprandial glucose concentrations was assessed in patients with non-insulin-dependent diabetes (NIDDM) and healthy volunteers. The study design consisted of a two-way crossover, single-dose administration of 10 g prehydrated high viscosity HPMC, or placebo, with a standard carbohydrate-rich meal. In patients with NIDDM, HPMC reduced blood glucose concentrations at the 60-, 75-, 90-, 120- and 150-min sampling intervals, with an average reduction in the maximum postprandial blood glucose concentration, Cmax, of 24% (P < 0.05). The time at which the maximum concentration was reached, Tmax, remained unchanged. The area under the blood concentration versus time plot, AUC0-6h, was reduced by an average of 15% (P < 0.05). The blood concentration profile of insulin followed that of glucose. Concentrations were significantly lower than in the placebo phase only at the 120-min sampling time, while pharmacokinetic parameters (Cmax, Tmax and AUC0-6h) were unchanged. These results suggest that alterations in the blood glucose profile are mediated by luminal events rather than by changes in hormonal response. In contrast to the NIDDM patients, neither the pharmacokinetic parameters nor the blood glucose concentrations at specific sampling times were significantly affected by the co-administration of HPMC in healthy volunteers. Overall, the results of this study suggest that HPMC may be a useful adjunct in the management of NIDDM.

Adult↗

The flexibility of low molecular weight double-stranded DNA as a function of length. I. Light scattering measurements and the estimation of persistence lengths from light scattering, sedimentation and viscosity.

In the preceding paper are described the isolation and physical characterization of seven narrowly disperse fractions of calf thymus DNA in the molecular weight range 0.3 to 1.3 X 10(6) daltons. Herein, we have determined by light scattering the molecular weights and root mean square radii of these fractions in a solvent comprising 0.2 M NaCl, 2 mM EDTA, 2mM Na-PO4,pH7. Measurements were made in a modified Wippler-Scheibling photometer to a 20 degree lower limit of scattering angle on solutions rendered virtually dust-free by procedures described. The optical anisotropies of the DNA fractions were measured permitting the experimental molecular weights and root mean square radii to be corrected to their true values. From these values, with appropriate polydispersity corrections, we calculate a Kratky-Porod persistence length, a, of 54.0 +/- 5.6 nm which is invariant over the molecular range examined. From the sedimentation coefficients (preceding paper) and the theory of Yamakawa and Fujii, we calculate a to be 66 nm, a value found to apply equally well to several DNA samples of various origins whose sedimentation rates are known in themolecular weight range from about 4 X 10(4) to 10(8) daltons. Similarly, from the intrinsic viscosities and the theory of Yamakawa and Fujii, we calculate a to be 59 nm, which again adequately applies to a number of DNA samples whose viscosities have been measured by other workers in the molecular wieght range 3 X 10(5) to 10(8) daltons. The Flory-Mandelkern paramerter, beta, was found to vary with molecular weight in the manner predicted by the theory of Yamakawa and Fujii. The average value of a from the three sets of measurements is 60 +/- 6nm, which we believe applies to double-stranded DNA molecules, independent of chain length, over the whole range of molecular weights from which reliable data exist.

Animals↗

Viscosity scaling and protein dynamics.

The rates of molecular motions in the interior of some proteins were found to scale with an inverse power of the external solvent viscosity. The data were explained by a flexible protein structure whose dynamics is partially controlled by the solvent. Reaction dynamics in the presence of structural fluctuations with finite lifetimes lead to a dynamic friction coefficient defined by a generalized Langevin equation and a fluctuation-dissipation theorem. A model for the dynamic friction is derived assuming that the fluctuation spectrum at the reaction site involves two components: solvent-independent diffusion of local structural defects in the protein matrix and global fluctuations coupled to the solvent. The theory is applied to the viscosity dependence of molecular oxygen-binding rates in sperm whale myoglobin.

Mathematics↗

The effect of viscosity on the accessibility of the single tryptophan in human serum albumin.

When reactions take place with one of the reactants tied to protein matrix, movements along the reaction coordinate towards the transition state can become coupled to structural fluctuations of the protein matrix. This investigation aims to test the assumptions underlying the arguments supporting such a coupling. A coupling is allowed only if the activation barrier is high and broad enough as shown to be the case for the proton catalyzed isotope exchange at Trp-63 of lysozyme. In the present investigation the activation barrier for the same reaction has been lowered radically in an effort to show that the coupling, as measured by the dependence of rate on solution viscosity, will diminish and ideally vanish, despite the unchanged effects of cosolvents on the chemical activities of all the reactants. The isotope exchange rate at the indole nitrogen of the single tryptophan residue of human serum albumin was measured with UV. This residue is rigidly held to the protein surface and the solvent access, although restricted, corresponds to a partially exposed residue. As a consequence, the isotope exchange rates and the bimolecular quenching rate of fluorescence by acrylamide, also measured, are high. The experiments were carried out at pH 5.2 where the molecule is in the N-form and the exchange is catalyzed by OH- ions. The activation energy of the hydroxyl catalyzed reaction is 22 kJ lower than for the proton catalyzed process. Under these conditions the exchange rate is viscosity independent both in the case of glycerol and in ethylene glycol.(ABSTRACT TRUNCATED AT 250 WORDS)

Deuterium↗

Cytoskeleton in the archaebacterium Thermoplasma acidophilum? Viscosity increase in soluble extracts.

Thermoplasma acidophilum has no cell wall, and so its irregular shape implies the presence of a cytoskeleton. When soluble extracts of T. acidophilum were incubated in vitro they increased in viscosity, suggestive of a polymerizable component. Optimal conditions for the viscosity increase coincided with physiological ionic concentrations. Electron micrographs of negatively stained extracts showed a meshlike lattice of elements 10 nm in diameter similar to nuclear lamins. However, immunologically there was no cross-reaction with lamins nor with the other eukaryotic cytoskeletal proteins tested: tubulin, calmodulin, giardin, actin or myosin.

Adenosine Triphosphate↗

Correlation of B coefficient of viscosity for monovalent salts with effects on binding of human follitropin to receptor.

Various monovalent salts have been shown to inhibit the binding of radioiodinated human follitropin [( 125I]hFSH) to receptors present in calf testis. We examined the effects of salts on the affinity constant (Ka) and number of binding sites (R0) in the FSH-calf testis receptor system. A constant amount of [125I]hFSH and increasing amounts of unlabeled hFSH were allowed to bind to a constant amount of receptor in the absence or presence (0.10 M) of halides, nitrates or acetates of the alkali ions at 20.0 degrees C. There was an appreciable variation of the affinity constant depending on the salt being used, but there was no change in receptor number (R0). Within an alkali cation series (e.g. NaCH3CO2, NaF, NaCl, NaBr, NaNO3, NaI or a similar potassium series) the affinity decreased with decreasing B coefficient of viscosity. Within a halide series (e.g. LiCl, NaCl, KCl, RbCl or the similar bromide series) the minimum value of the affinity constant occurred with the Na+ salt. The special inhibitory potency of Na+ may suggest that it has a unique interaction with calf testis FSH receptor or with follitropin. The studies indicate a correlation between the B coefficient of viscosity and the ability of salts to inhibit binding of [125I]hFSH to receptor.

Acetates↗

Sulfapyridine and sulfones decrease glycosaminoglycans viscosity in dermatitis herpetiformis, ulcerative colitis, and pyoderma gangrenosum.

Shortly after the introduction of sulfa drugs, sulfapyridine was found to have unique therapeutic properties, unrelated to antibacterial activity. Later, sulfones were found to share the same properties. The disorders initially improved were dermatitis herpetiformis, pyoderma gangrenosum, subcorneal pustular dermatosis, acrodermatitis continua, impetigo herpetiformis and ulcerative colitis. They were also sometimes helpful in many other disorders. They are effective in select disorders characterized by edema followed by granulocytic inflammation or edema followed by vesicle or bullae formation. The sulfones work in low doses in leprosy and their mode of action is not fully understood. Several pieces of experimental information are available. It is proposed that these drugs are entering or influencing the protein moiety of glycosaminoglycans and decreasing tissue viscosity. This decreased tissue viscosity prevents edema and dilution of tissue fluid and decreases acute inflammation and vesicle and bullae formation.

Clofazimine↗

High viscosity of newborn extracellular matrix is the etiology of erythema toxicum neonatorum: neonatal jaundice?: hyaline membrane disease?

At the time of birth, the fetal ground substance is under the influence of maternal and placental hormones. Hormones are known to exert significant effect on ground substance. The ground substance viscosity that is ideal for intrauterine life is too viscous for the newborn. Sites of minor skin trauma develop swelling, inflammation and pustules. Dilution of tissue fluids causes formation of some of the mediators of inflammation including those responsible for intraepidermal pustule formation. The newborn responds to inflammation with eosinophilic granulocyte. The clinical lesions referred to as erythema toxicum neonatorum are known to localize at pressure sites. The skin lesions correct as maternal and placental hormone influence weakens. The entity occurs in 31 to 72% of white infants. The hormone induced viscosity changes may aid in better understanding some aspects of neonatal physiology; such as jaundice of the neonatal period, the eosinophilic granulocyte inflammatory response of the neonatal period and hyaline membrane disease.

Erythema↗