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Biomedical subjects

I F Miller

Publications and source records attributed to I F Miller.

At least 37 records · Page 2Linked to original sources

Regulation of ciliary beat frequency by autonomic mechanisms: in vitro.

The ciliated epithelium of the mammalian trachea separates the neurohumoral milieu of the tissue from that of the environment of the airway lumen. To determine whether specific autonomic receptors regulating ciliary beat frequency (CBF) were located on mucosal or serosal sides, we measured CBF by heterodyne mode correlation analysis laser light scattering in bovine tracheal tissues mounted in a two-sided chamber. A beta 2-adrenergic agonist, fenoterol, at 10(-7) M, stimulated serosal CBF from 7.9 +/- 1.3 to 20.2 +/- 5.8 Hz (P less than 0.01) and mucosal CBF from 6.6 +/- 0.9 to 14.7 +/- 4.6 Hz (P less than 0.01). A muscarinic cholinergic agonist, methacholine, at 10(-7) M, increased mucosal CBF from 8.4 +/- 1.0 to 19.5 +/- 5.5 Hz (P less than 0.01) and serosal CBF from 8.0 +/- 0.9 to 15.4 +/- 5.0 Hz (P less than 0.01). The differences in stimulation of CBF on the mucosal and serosal sides between fenoterol and methacholine were significant (P less than 0.01). Studies in which these autonomic agonist stimulating effects were inhibited by their respective antagonists, propranolol and atropine sulfate, demonstrated that CBF can be regulated independently by mediators both in the submucosa and within the mucus lining.

Animals↗

Hemodynamic effects of exchange transfusions with liposome-encapsulated hemoglobin.

The ability of liposome-encapsulated hemoglobin (LEH) to sustain life in rats at hematocrits 45% below lethal levels was demonstrated by virtually total exchange transfusions. All control animals (transfused with 7% albumin in Erilyte) died at a mean hematocrit of 5.35%, with corresponding oxygen-carrying capacity of 2.65 ml/100 ml. All LEH animals survived with a final RBC hematocrit of 2.96% and an oxygen-carrying capacity of 7.05 ml/100 ml. LEH animals maintained normal blood pressures and essentially normal cardiac output, while control animals failed to do so. Systemic vascular resistance of control animals at 78% exchange was reduced to 33% of baseline, while of LEH animals at 92% exchange, to 80% of baseline. In-vitro measurements were made of LEH and RBC suspension viscosity as a function of shear rate, suspension concentration, and lipid membrane composition. The viscous behavior of the LEH suspensions was non-Newtonian and nearly the same as suspensions of natural RBC prepared similarly, though LEH suspensions had slightly higher viscosity.

Animals↗

Cardiorespiratory effects of exchange transfusions with synthetic erythrocytes in rats.

Synthetic erythrocytes (SE) can maintain life with near-normal hemodynamics in rats with hematocrits (Hct) 45% below the lethal levels, as shown when we exchanged virtually all their blood. We compared cardiorespiratory variables between the SE and control animals, which were hemodiluted with 7% albumin, at the 78% exchange level. In addition, SE animals at 92% exchange were compared with the 78% exchange and baseline levels. All our control rats died at Hct above 5% with corresponding oxygen-carrying capacity of 2.65 ml/dl. All SE rats survived, having a final Hct of 2.96% with an oxygen-carrying capacity of 7.05 ml/dl. SE animals maintained normal BP and marginally increased cardiac output, while control animals did not. Vascular resistance of control animals at 78% exchange was reduced to 30%, while in SE animals at 92% exchange, vascular resistance was lowered to 80% of baseline values.

Animals↗

Measurements of viscosity of synthetic erythrocyte suspensions.

Measurements were made of the viscosity of suspensions of synthetic erythrocytes composed of hemoglobin solutions encapsulated in liposomes, as a function of shear rate, temperature, suspension concentration, lipid membrane composition, and the viscosity of the suspending medium. It was found that the viscous behavior of the synthetic erythrocyte suspensions was non-Newtonian and nearly the same as that of suspensions of natural erythrocytes prepared similarly, with the major difference being that synthetic erythrocyte suspensions are somewhat more viscous. Suspensions of Fluosol FC-43 prepared similarly were found to be essentially Newtonian fluids, and substantially different and more viscous than either erythrocyte suspension. The higher viscosity of synthetic erythrocyte suspensions probably accounts for the ability of these suspensions to maintain normal systemic vascular resistance in transfusion experiments, in spite of the fact that synthetic erythrocytes are smaller than natural erythrocytes.

Blood Proteins↗

Automated measurement of ciliary beat frequency.

Measurements of ciliary beat frequency using video images are dependent on observer interpretation. To obtain objective estimates of ciliary beat frequency from video-image sequences, a computer-based method was developed. Regions of interest of video-image sequences were selected and digitized. Variations in numerical values representing light intensity resulting from cilia beating were extracted and analyzed using autocorrelation techniques. The ciliary beat frequencies obtained for 14 in vitro experiments on ciliated cells or epithelium from the frog palate (Rana catesbeiana) over the range of frequencies 2-25 Hz correlated well with independent observer measurements (r = 0.979). The addition of such computer-based methods to video observer-based systems allows more objective and efficient determinations of ciliary beat frequency.

Animals↗

Effects of lipid-soluble substances on the thermotropic properties of liposome filtration.

Filtration of dimyristoyl phosphatidylcholine or dipalmitoyl phosphatidylcholine liposomes at various temperatures from 3 to 60 degrees C revealed a discontinuous change in filtration behavior centered about the gel-to-liquid crystal transition temperature. This change was continuous at temperatures immediately above or below the transition temperature. Although pure dipalmitoyl phosphatidylcholine liposomes are in the gel state at 22 degrees C, passage of liposomes composed of dipalmitoyl phosphatidylcholine and cholesterol through the filters at 22 degrees C gave results similar to those obtained with liquid-crystal liposomes. Low cholesterol concentrations were nearly as effective as high concentrations in producing this behavior; this observation is consistent with a shear mechanism for reduction of liposome size, since the stress induced by passage of the otherwise rigid liposome through a small pore would be relieved by fracture at a lattice imperfection. Liposomes composed of egg phosphatidylcholine and cholesterol were retained by the filters to a slightly greater extent than pure egg phosphatidylcholine liposomes; these results are consistent with the known condensing effect of cholesterol on liquid-crystal lipsomes and a shear mechanism occurring with filtration. Liposomes composed of dipalmitoyl phosphatidylcholine and either dipalmitoyl phosphatidic acid or dicetyl phosphate were filtered at 22 degrees C; they showed a filtration characteristic similar to liquid-crystal liposomes. Inclusion of the water-soluble dyes eosin Y or Evans blue in dipalmitoyl phosphatidylcholine liposomes resulted in filtration at 22 degrees C which was similar to that observed for liquid-crystal liposomes. The dyes, sodium fluorescein, 6-carboxyfluorescein and fluoresceinisothiocyanate dextran, did not alter 22 degrees C liposome filtration.

Cholesterol↗

Liposome filtration. Dependence on transition temperature.

Liposomes formed by vortexing and passed through polycarbonate surface retention membranes showed appreciable differences in filtration behavior depending on the temperature of filtration relation relative to the liposome gel-liquid crystal transition temperature. Below transition, liposomes were filterable and size distributions could be determined; the cumulative volume distributions were log-normal. Above transition, liposomes were not filterable: smaller liposomes were formed until a limiting size was reached. These results suggest that liquid crystal liposome size distributions cannot be determined by filtration. This filtration behavior is a physical property of liposomes, related to the gel-liquid crystal transition, not previously reported. This property could be exploited as a new method for controlling liposome size distributions, but the implications for lipid membranes, including biological membranes, are general.

Filtration↗

Synthetic erythrocytes from lipid encapsulated hemoglobin.

Synthetic erythrocytes have been formed by encapsulating concentrated hemoglobin solutions in microcapsules consisting of phospholipid-cholesterol mixtures. The microcapsules (hemosomes) are of the order of one micron in diameter or smaller, are osmometers, are somewhat sturdier than erythrocytes (RBC), and are stable upon freezing. They have essentially the same oxygen and carbon dioxide carrying capacity as RBC, and can be tailored to have the same electrokinetic properties. They appear to evoke no immune response in the rat. In experiments involving complete replacement of natural blood hemosomes appear to sustain life. Rats can live and breathe normally without signs of hypoxia or acidosis for extended periods of time after the hematocrit of natural RBC drops well below the critical level. Synthetic erythrocytes do not appear to cause disseminated coagulation, microembolism, or any observable changes in internal organs.

Animals↗

Kinetic studies of a doubly bound red cell antigen-antibody system.

The Polybrene method for detection of red cell antibodies which utilizes continuous flow equipment was modified so that kinetic studies could be performed on red cell antibodies doubly bound between adjacent red cells. In the anti-Rh(o)-Rh(o) erythrocyte system, deaggregation by temperature was studied over an antibody concentration range of from approximately 1 to 500 antibody molecules per erythrocyte, a residence time range of approximately eightfold, and a temperature range of from 10 to 55 degrees C. The rate of dissociation of antigen-antibody complex, as determined from deaggregation of antibody-dependent red cell aggregates, was found to be of apparent zero order. The apparent activation energy for the antigen-antibody reaction under the experimental conditions was determined and found to be higher than would be expected for singly bound antigen-antibody systems. Possible explanations are considered for these findings in terms of an antigen-antibody bond-breaking model.

Antigen-Antibody Complex↗