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Biomedical subjects

K Arnold

Publications and source records attributed to K Arnold.

At least 19 recordsLinked to original sources

The influence of dextran sulfate on influenza A virus fusion with erythrocyte membranes.

Dextran sulfate suppresses the low pH-induced fusion of influenza virus A/Brazil 11/78 with erythrocyte membranes, as shown by fluorescence dequenching assay, using the fluorophore octadecylrhodamine B chloride (R18). Inhibition of fusion was maximal at pH 5.0, while at higher pH values (> 5.6) fusion was not affected. Hemolysis of intact red blood cells by influenza A virus at low pH values is also prevented by dextran sulfate. The inhibiting effect of the polymer is mainly ascribed to repression of virus attachment. Evidence is given that the conformational change of the virus envelope protein hemagglutinin (HA) responsible for triggering fusion is not affected by the polymer.

Anilino Naphthalenesulfonates

Randomised trial of artesunate and mefloquine alone and in sequence for acute uncomplicated falciparum malaria.

The increasing frequency of therapeutic failures in falciparum malaria in Thailand shows an urgent need for effective drugs or drug combinations. Artesunate, a qinghaosu derivative, is effective in clearing parasitaemia rapidly, but the recrudescence rate can be as high as 50%. We have compared artesunate followed by mefloquine with each drug alone in acute, uncomplicated falciparum malaria. 127 patients were randomly assigned treatment with artesunate (600 mg over 5 days), mefloquine (750 mg then 500 mg 6 h later), or artesunate followed by mefloquine. All patients were admitted to hospital for 28 days to exclude reinfection. Cure was defined as no recrudescence during the 28 days' follow-up. The cure rates for mefloquine and artesunate alone were 81% (30/37 patients) and 88% (35/40); the combination was effective in all of 39 patients. Fever and parasite clearance times were significantly shorter in the groups that received artesunate than in the mefloquine-only group. The frequency of nausea and vomiting was slightly, but not significantly, higher among patients who received both drugs than in the other groups. The combination of artesunate followed by mefloquine is highly effective and well tolerated in patients with acute, uncomplicated falciparum malaria in Thailand.

Acute Disease

Association of lysozyme to phospholipid surfaces and vesicle fusion.

Lysozyme-induced fusion of phosphatidylserine (PS) vesicles was studied as a function of pH. Fusion, monitored by lipid-mixing, was measured by following the dilution of pyrene-labelled phosphatidylcholine, incorporated in PS vesicles, into unlabelled bilayers. It is demonstrated that lysozyme-induced fusion is pH-dependent and significant fusion is triggered at pH 5 or below. The interaction of lysozyme with the vesicle bilayer was characterized by measuring resonance energy transfer from tryptophane, present in the protein, to pyrene. It is shown that concomitant with fusion, a strong resonance energy transfer signal appears at pH 5 or below. Furthermore, in monolayer experiments it was found that addition of lysozyme to the subphase caused an increase in surface pressure, when the pH was kept below 5.5. Very low concentrations of lysozyme sufficed to bring about the observed effects. The results are taken to indicate that lysozyme-induced fusion results from penetration of protein into the hydrophobic core of the bilayer, occurring at acidic pH.

Energy Transfer

Motor control prior to movement onset: preparatory mechanisms for pointing at visual targets.

The present study investigated the mechanisms involved in the preparation of pointing movements in humans. We provided visual precues on the location of the upcoming target, and registered the effect of these precues on the reaction time (RT = interval between target appearance and movement onset). Generally, precues were found to reduce RT, suggesting that some aspects of the preparatory process have been advanced in time. In Exp. 1, precues fully specified the direction required for the upcoming movement while indicating only a range of movement amplitudes; in Exp. 2, precues fully specified the amplitude and indicated a range of directions. In both experiments, RT was shorter than in control trials without precues, and gradually increased with the size of the precued amplitude or direction range. This result suggests that the preparation of either parameter is possible without knowing the precise value of the other, i.e. amplitude and direction are not prepared in a fixed order. Furthermore, our results are consistent with the view that movement preparation includes a progressive contraction of the precued range towards the final value. The speed of this process can be estimated as 0.31 cm/ms for amplitude, and 1.7 deg/ms for direction ranges. In Exp. 3 and 4, precues indicated both amplitude and direction as ranges only. The size of the amplitude range was held constant while the size of the direction range was varied (Exp. 3), or vice versa (Exp. 4). Under these conditions, RT increased with the size of the varied range.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Dextran sulfate-dependent fusion of liposomes containing cationic stearylamine.

The incorporation of the positively charged stearylamine into phosphatidylcholine liposomes was studied by measuring electrophoretic mobilities. Up to a molar ratio SA/PC = 0.5 an increase of the positive zeta potential can be observed. Addition of the negatively charged macromolecule dextran sulfate leads to a change of the sign of the surface potential of the PC/SA liposomes indicating binding of the macromolecule to the surface. This process is accompanied by an increase in turbidity, which is dependent on the molecular weight of the dextran sulfate and the SA concentration (measured by turbidimetry). Using the NBD/Rh and Pyr-PC fluorescence assays the fusion of SA containing liposomes was investigated. A strong influence of the SA content and molecular weight of dextran sulfate on the fusion extent was observed. The fusion extent is proportional to the SA content in the PC membrane and the molecular weight of dextran sulfate. PC/SA/PE liposomes exhibit a higher fusion extent after addition of dextran sulfate compared to PC/SA liposomes indicating that PE additionally destabilizes the bilayer. Freeze-fracture electron microscopy reveals that the reaction products are large complexes composed of multilamellar stacks of tightly packed, straight membranes and aggregated vesicles. The tight packing of the membranes in the stacks (and the narrow contact of the aggregated vesicles) indicates a strong adherence of opposite membrane surfaces induced by dextran sulfate.

Amines

Effect of anionic polymers on fusion of Sendai virus with human erythrocyte ghosts.

The effect of anionic polymers (dextran sulfate, heparin and chondroitin sulfate) on fusion of Sendai virus with erythrocyte ghosts was studied. The effect of pH on the activity of these anionic polymers was also investigated. In order to examine the interaction of such polymers with the Sendai virion and erythrocyte ghost surfaces, the binding of virions to erythrocyte ghosts and the aggregation of virions and/or erythrocyte ghosts were also measured with respect to the same parameters. It was found that the anionic polymers suppressed the fusion of Sendai virus with erythrocyte ghosts. The order of effectiveness of the polymers in suppression was dextran sulfate greater than heparin greater than chondroitin sulfate, for the application of a same quantity (weight/ml) of the polymers. The lower the pH of the suspending medium, the more effective were the polymers in suppressing virion-erythrocyte ghost aggregation and fusion. The suppression of fusion was dependent on the concentration of the polymers applied: the higher the concentration of the polymer applied, the more the suppression was observed. Evidence from binding studies, turbidity measurements and electrophoretic mobility measurements indicates that the anionic polymers interact preferentially with the virion surface.

Adsorption

Role of functional groups of human plasma and luminol in scavenging of NaOCl and neutrophil-derived hypochlorous acid.

Hypochlorous acid HOCl/OCl- and other oxidants derived from stimulated polymorphonuclear leukocytes are involved in tissue damage during a number of pathological processes. In order to obtain more detailed information on possible reactions of HOCl/OCl- the effects of both NaOCl and PMN-derived hypochlorous acid on functional groups of amino acid solutions and human plasma are studied. In valine and lysine solutions NaOCl diminishes the number of amino groups in a molar ratio of 1:1 between NaOCl and amino groups. In cysteine and methionine samples the decrease of amino groups starts only after all sulfhydryl or thioether groups are oxidized by NaOCl. If freshly prepared human plasma is treated with increasing amounts of NaOCl all plasma SH groups are oxidized first, then probably the thioether groups and only after this the amino groups are affected. Furthermore, it was found, that the reactivity of luminol against NaOCl is similar to that of amino groups. Increasing amounts of SH groups of components of human plasma are oxidized by incubation with PMA-stimulated polymorphonuclear leukocytes dependent on the incubation time. Plasma amino groups are not affected under the same experimental conditions. The addition of plasma to FMLP-stimulated PMN in the presence of luminol decreases that part of chemiluminescence caused by extracellularly generated hypochlorous acid. Plasma samples pretreated with NaOCl cause a lower inhibition of light generation in FMLP-stimulated PMN only when more than 4.10(-8) mol NaOCl per mg protein are used to pretreat plasma. It is assumed that in the development of tissue injuries caused by infiltrated PMN the following sequence of damage occurs in accessible tissue regions. First, the sulfhydryl groups are oxidized, then the thioether groups, and only after this amino and other target groups are affected.

Amino Acids

Immune-mediated hemolytic anemia in a horse.

An 18-year-old Quarter Horse gelding was determined to have immune-mediated hemolytic anemia after detection of autoagglutination of RBC spherocytosis as well as a positive direct Coombs test result. A lack of response to treatment with corticosteroids necessiated the administration of cyclophosphamide and azathioprine. The anemia resolved after treatment with chemotherapeutic drugs.

Adrenal Cortex Hormones

The influence of particle size and multiple apoprotein E-receptor interactions on the endocytic targeting of beta-VLDL in mouse peritoneal macrophages.

Low density lipoprotein (LDL) and beta-very low density lipoprotein (beta-VLDL) are internalized by the same receptor in mouse peritoneal macrophages and yet their endocytic patterns differ; beta-VLDL is targeted to both widely distributed and perinuclear vesicles, whereas LDL is targeted almost entirely to perinuclear lysosomes. This endocytic divergence may have important metabolic consequences since beta-VLDL is catabolized slower than LDL and is a more potent stimulator of acyl-CoA/cholesterol acyl transferase (ACAT) than LDL. The goal of this study was to explore the determinants of beta-VLDL responsible for its pattern of endocytic targeting. Fluorescence microscopy experiments revealed that large, intestinally derived, apoprotein (Apo) E-rich beta-VLDL was targeted mostly to widely distributed vesicles, whereas small, hepatically derived beta-VLDL was targeted more centrally (like LDL). Furthermore, the large beta-VLDL had a higher ACAT-stimulatory potential than the smaller beta-VLDL. The basis for these differences was not due to fundamental differences in the means of uptake; both large and small beta-VLDL were internalized by receptor-mediated endocytosis (i.e., not phagocytosis) involving the interaction of Apo E of the beta-VLDL with the macrophage LDL receptor. However, large beta-VLDL was much more resistant to acid-mediated release from LDL receptors than small beta-VLDL. Furthermore, partial neutralization of the multiple Apo Es on these particles by immunotitration resulted in a more perinuclear endocytic pattern, a lower ACAT-stimulatory potential, and an increased sensitivity to acid-mediated receptor release. These data are consistent with the hypothesis that the interaction of the multivalent Apo Es of large beta-VLDL with multiple macrophage LDL receptors leads to a diminished or retarded release of the beta-VLDL from its receptor in the acidic sorting endosome which, in turn, may lead to the widely distributed endocytic pattern of large beta-VLDL. These findings may represent a physiologically relevant example of a previously described laboratory phenomenon whereby receptor cross-linking by multivalent ligands leads to a change in receptor targeting.

Animals

Effect of dextran sulfate on fusion of Sendai virus with human erythrocyte ghosts.

The effect of dextran sulfate on the fusion of Sendai virus and erythrocyte ghosts was studied as a function of dextran sulfate concentration and pH of cell suspension solutions. In order to examine the interaction of dextran sulfate with Sendai virion and erythrocyte ghost surfaces, the turbidity of cell suspensions was also measured with respect to the same parameters as above. It was found that dextran sulfate inhibited the fusion of Sendai virus with erythrocyte ghosts. The lower the pH of the suspension solution was, the more effective was dextran sulfate in suppressing virion erythrocyte ghost fusion. Dextran sulfate of a higher molecular weight was slightly more effective in suppressing fusion than that of a lower molecular weight. From turbidity measurements of Sendai virus and erythrocyte ghost suspensions, it is likely that dextran sulfate interacts preferentially with Sendai virion surfaces and inhibits interaction between Sendai virions and erythrocyte ghosts.

Depression, Chemical

Partition of serum lipoproteins in a polyethylene glycol/dextran two-phase system.

Aqueous two-phase systems containing polyethylene glycol (PEG) and dextran as phase forming polymers were used for the partition of unmodified and hypochlorite modified lipoproteins. Low density lipoprotein (LDL) was separated from high density lipoprotein (HDL) by sequential ultracentrifugation from human plasma. In agreement with the higher electrophoretic mobility, high density lipoprotein shows a higher value of the partition coefficient in contrast to low density lipoprotein. An increase in the concentration of chloride ions reduces the enrichment of lipoprotein in the top phase and favours the accumulation of aggregated material at the interface. The partition coefficient strongly depends on the age of the lipoprotein sample. Differences in the value of the partition coefficient could be obtained for the lipoprotein fractions HDL-2 and HDL-3. Hypochlorite modified LDL shows higher values of the partition coefficient due to the higher negative charge of the modified lipoprotein particle.

Chemical Phenomena

Modification of low density lipoproteins by sodium hypochlorite.

Human low density lipoproteins (LDL) were incubated with increasing amounts of sodium hypochlorite. A decrease of the number of free amino groups on the LDL surface starts only upon addition of 30-40 moles NaOCl per mole apoB, whereas all detectable SH groups are oxidized after addition of nearly 17-20 moles NaOCl. All hypochlorite-modified LDL samples have a higher electronegative surface charge compared with native LDL as revealed by agarose gel electrophoresis and partition of LDL in an aqueous polyethylene glycol/dextran two-phase system. The more NaOCl is used to alter LDL, the higher is the electrical surface charge. Changes in surface charge are found already at low NaOCl concentrations where no decrease of amino groups is detected. It is assumed that changes in surface charge are caused by the formation of monochloramines and especially at low degrees of modification by a further unknown contribution. An effect on the primary structure of apoB or peroxidation-like changes in NaOCl-altered LDL could not be found under our experimental conditions. The results are discussed with respect to such modifications under in vivo conditions by hypochlorous acid generated in stimulated phagocytosing cells.

Dextrans

Aggregation of human plasma high density lipoproteins induced by poly(ethylene glycol).

The influence of different surface charge densities (induced by varying pH, addition of positively charged amphiphilic molecules and chemical modification) of high density lipoproteins (isolated by ultracentrifugation) on poly(ethylene glycol) induced aggregation was studied. The effects of different molecular masses of PEG, HDL concentration and the presence of other serum proteins on the PEG mediated aggregation were investigated. The PEG concentration necessary for HDL aggregation is inversely proportional to the used HDL concentration and its molecular weight, and is directly proportional to the presence of other proteins and the magnitude of the negative surface charge density of HDL. The results are in accordance with the predictions of the volume exclusion theory taking into account the influence of repulsive electrostatic forces on the interaction of HDL particles.

Chemical Precipitation

Surface dielectric constant, surface hydrophobicity and membrane fusion.

Membrane fusion induced by ions and its associated membrane property, surface dielectric constant, were studied with the use of acidic and neutral phospholipid vesicles. The fusion of vesicles was monitored by utilizing two fluorescence fusion assays: fluorescence content mixing method and fluorescence labelled membrane component dilution method. For the surface dielectric constant measurements, a fluorescence method was used which detected the environmental effect on the membrane surface upon the addition of various fusogenic cations. Also, the effects of poly-(ethylene glycol) on both fusion and surface dielectric properties were examined. It was found that the extent of fusion correlated well with the degree of lowering in the dielectric constant of the surface membrane, which corresponds to the increase in hydrophobicity of the membrane surface. This agrees with the previously obtained experimental results that the increase in interfacial tension of the membrane, which also corresponds to the increase in surface hydrophobicity, correlates with the extent of membrane fusion.

Calcium

[Biophysical research on condylar temporomandibular joint growth].

On the basis of results obtained from the study of mechanically induced potentials and pulsed 1H-NMR investigations of the condylar cartilage of the TMJ in vitro, the hypothesis is advocated that the internal regulation of condylar growth takes place with the aid of bioelectric signals. Reference in made to the importance of the studies for a future noninvasive diagnosis of condylar growth in the course of functional orthopedic treatment, using "dynamic" magnetic resonance imaging.

Animals

Depletion flocculation and depletion stabilization of erythrocytes.

At dextran (Mw approximately 500,000) concentrations from 2 to approximately 10%, suspensions of normal human erythrocytes flocculate in small convex agglutinates. At dextran concentrations greater than 10%, the erythrocytes resegregate in a stable monodisperse suspension. At all these dextran concentrations, the erythrocytes are coated with considerable amounts of dextran. It can be argued that at dextran concentrations from 2 to 10%, as well as at dextran concentrations greater than 10%, there is a thin layer, which is depleted of dextran, between the dextran layer adsorbed onto the erythrocytes and the bulk dextran solution. It can also be shown that there is a repulsive interaction between the two layers of dextran: one adsorbed and one free. When the adsorbed dextran layer is the most concentrated, stability must ensue, and when the dextran in free solution is the most concentrated, flocculation should occur. Below 7% dextran, the concentration of free dextran is higher than the adsorbed concentration; above 10% dextran that situation is reversed. These data correlate well with the depletion flocculation predicted for the lower concentration and the depletion stabilization predicted for the higher dextran concentration.

Biophysical Phenomena