Basic properties of beta adrenergic receptors mediating melanosome dispersion of melanophores in a cyprinid fish, Zacco temmincki

In melanophores of a cyprinid fish, Zacco temmincki, receptor mechanisms of melanosome dispersion induced by catecholamines were examined. While possessing a melanosome-aggregating action in higher concentrations, isoproterenol and epinephrine in lower concentrations acted to disperse melanosomes. Norepinephrine, epinine and dopamine altered their action from melanosome aggregation to melanosome dispersion after alpha adrenergic blockade. The catecholamine-induced melanosome dispersion was inhibited by beta adrenergic blocking agents. Mediation of dispersion is regulated through beta adrenergic receptors. The beta adrenergic responses were unaffected by mersalyl, a sulfhydryl inhibitor. A prospective substance acting in dispersing melanosomes appears to be adrenaline, but not noradrenaline.     

Pigment movements in fish melanophores: Morphological and physiological studies

Summary Cyclic adenosine monophosphate (cAMP) has been shown to cause pigment dispersion in amphibian and fish melanophores. Since pigment displacements in melanophores of Pterophyllum scalare are known to be accompanied by assembly and disassembly of microtubules, the effect of cAMP on this process was investigated. Melanophores of isolated scales were treated with cAMP in the presence of vinblastine, a potent antimicrotubular agent. During the initial phase of vinblastine action, cAMP as well as its dibutyryl derivative are capable of counteracting the inhibitory effects of vinblastine on pigment dispersion. In addition, cAMP retains the velocity of pigment dispersion at about the maximum level during 1 hour experiments. Pigment aggregation was unaffected by cAMP. Since pigment dispersion in Pterophyllum-melanophores is accompanied by assembly of microtubules, it is concluded that cAMP influences, at least in part, melanosome dispersion through facilitation of microtubule assembly.     

Fast and accurate fitting of relaxation dispersion data using the flexible software package GLOVE

Relaxation dispersion spectroscopy is one of the most widely used techniques for the analysis of protein dynamics. To obtain a detailed understanding of the protein function from the view point of dynamics, it is essential to fit relaxation dispersion data accurately. The grid search method is commonly used for relaxation dispersion curve fits, but it does not always find the global minimum that provides the best-fit parameter set. Also, the fitting quality does not always improve with increase of the grid size although the computational time becomes longer. This is because relaxation dispersion curve fitting suffers from a local minimum problem, which is a general problem in non-linear least squares curve fitting. Therefore, in order to fit relaxation dispersion data rapidly and accurately, we developed a new fitting program called GLOVE that minimizes global and local parameters alternately, and incorporates a Monte-Carlo minimization method that enables fitting parameters to pass through local minima with low computational cost. GLOVE also implements a random search method, which sets up initial parameter values randomly within user-defined ranges. We demonstrate here that the combined use of the three methods can find the global minimum more rapidly and more accurately than grid search alone.     

Transit time dispersion in the pulmonary arterial tree

Knowledge of the contributions of arterialand venous transit time dispersion to the pulmonary vascular transittime distribution is important for understanding lung function and forinterpreting various kinds of data containing information aboutpulmonary function. Thus, to determine the dispersion of blood transittimes occurring within the pulmonary arterial and venous trees, imagesof a bolus of contrast medium passing through the vasculature ofpump-perfused dog lung lobes were acquired by using an X-ray microfocalangiography system. Time-absorbance curves from the lobar artery andvein and from selected locations within the intrapulmonary arterial tree were measured from the images. Overall dispersion within the lunglobe was determined from the difference in the first and second moments(mean transit time and variance, respectively) of the inlet arterialand outlet venous time-absorbance curves. Moments at selected locationswithin the arterial tree were also calculated and compared with thoseof the lobar artery curve. Transit times for the arterial pathwaysupstream from the smallest measured arteries (200-µm diameter) wereless than ~20% of the total lung lobe mean transit time. Transittime variance among these arterial pathways (interpathway dispersion)was less than ~5% of the total variance imparted on the bolus as itpassed through the lung lobe. On average, the dispersion that occurredalong a given pathway (intrapathway dispersion) was negligible. Similar results were obtained for the venous tree. Taken together, the resultssuggest that most of the variation in transit time in theintrapulmonary vasculature occurs within the pulmonary capillary bedrather than in conducting arteries or veins.

     

Passive Electrical Properties of Microorganisms: V. Low-Frequency Dielectric Dispersion of Bacteria

Very high dielectric constants have been observed for bacteria at low frequencies. High dielectric constants such as these can be explained by a theory which has been developed for the low-frequency dielectric dispersion of porous charged particles and which has been tested successfully through measurements with ion exchange resins. The bacterial cell wall is electrically similar to an ion exchange resin. Observations show that the theory provides a quantitative explanation for the low-frequency dielectric dispersion of bacteria.     

Iontophoretic release of cyclic AMP and dispersion of melanosomes within a single melanophore

Selective dispersion of melanosomes was often observed after iontophoretic injection of cyclic adenosine monophosphate (AMP) from a glass microelectrode positioned in a target melanophore in frog skin (as viewed from above through a microscope), with other melanophores in the field serving as controls. Because the skin has orderly arrays of several types of closely spaced cells, it is probable that at times the microelectrode also impales cells other than melanophores. When cyclic AMP injection inside a cell resulted in dispersion of melanosomes from a perinuclear position into dendritic processes, the onset of dispersion was relatively rapid, in many cases less than 4 min (mean time of onset, 5.3 +/- 2.9 [SD] min). A much slower dispersion (mean time of onset, 19.0 +/- 5.0 min) of melanosomes was observed when the microelectrode was positioned adjacent to a melanophore, and much larger quantities of cyclic AMP were released. In addition, no changes were observed for injections of 5'-AMP or cyclic guanosine monophosphate (GMP) through electrodes positioned inside or adjacent to melanophores. Potential measurements showed that after impaling a clell, a constant transmembrane potential could often be recorded over many minutes, indicating that the membrane tends to seal around the microelectrode. The results indicate that cyclic AMP acts more rapidly on the inside of a cell than when applied outside a cell and allowed to diffuse through the plasma membrane. This study introduces a model system whereby the properties of the plasma membrane and melanocyte-stimulating hormone (MSH) receptors can be studies within a single target cell.     

Residence time distributions of gas flowing through rotating drum bioreactors

Residence time distribution studies of gas through a rotating drum bioreactor for solid-state fermentation were performed using carbon monoxide as a tracer gas. The exit concentration as a function of time differed considerably from profiles expected for plug flow, plug flow with axial dispersion, and continuous stirred tank reactor (CSTR) models. The data were then fitted by least-squares analysis to mathematical models describing a central plug flow region surrounded by either one dead region (a three-parameter model) or two dead regions (a five-parameter model). Model parameters were the dispersion coefficient in the central plug flow region, the volumes of the dead regions, and the exchange rates between the different regions. The superficial velocity of the gas through the reactor has a large effect on parameter values. Increased superficial velocity tends to decrease dead region volumes, interregion transfer rates, and axial dispersion. The significant deviation from CSTR, plug flow, and plug flow with axial dispersion of the residence time distribution of gas within small-scale reactors can lead to underestimation of the calculation of mass and heat transfer coefficients and hence has implications for reactor design and scale-up.     

The dipolar origin of protein relaxation

1. A set of parameters is proposed to check the interpretation of the dielectric behaviour of protein solutions as a rigid-dipole relaxation of prolate ellipsoids of revolution in the frequency range between 20 kHz and 10 MHz. Besides the delta(b)-function of Scheraga, another analogous function (delta(a)) is presented to establish size and shape of globular proteins. A study of the influence of solvent viscosity on the dielectric dispersion also gives strong evidence in favour of rigid-dipole relaxation. 2. Measurements of the dielectric dispersion of monomer solutions of bovine serum albumin and transferrin are reported. Monomers of bovine serum albumin were obtained by fractionation on Sephadex G-150. Low-conductivity solutions of both proteins are obtained by passage through an ion-exchange resin. 3. Computer analysis of the experimental dispersion curves by use of a two-term Debye dispersion gives valuable information about transferrin and leads to an axial ratio 4.5 for a prolate ellipsoid of revolution. The dielectric increment of bovine serum albumin is very low and no conclusive results have yet been obtained.     

Myosin II and Rho kinase activity are required for melanosome aggregation in fish retinal pigment epithelial cells

In the retinal pigment epithelium (RPE) of fish, melanosomes (pigment granules) migrate long distances through the cell body into apical projections in the light, and aggregate back into the cell body in the dark. RPE cells can be isolated from the eye, dissociated, and cultured as single cells in vitro. Treatment of isolated RPE cells with cAMP or the phosphatase inhibitor, okadaic acid (OA), stimulates melanosome aggregation, while cAMP or OA washout in the presence of dopamine triggers dispersion. Previous studies have shown that actin filaments are both necessary and sufficient for aggregation and dispersion of melanosomes within apical projections of isolated RPE. The role of myosin II in melanosome motility was investigated using the myosin II inhibitor, blebbistatin, and a specific rho kinase (ROCK) inhibitor, H-1152. Blebbistatin and H-1152 partially blocked melanosome aggregation triggered by cAMP in dissociated, isolated RPE cells and isolated sheets of RPE. In contrast, neither drug affected melanosome dispersion. In cells exposed to either blebbistatin or H-1152, then triggered to aggregate using OA, melanosome aggregation was completely inhibited. These results demonstrate that (1) melanosome aggregation and dispersion occur through different, actin-dependent mechanisms; (2) myosin II and ROCK activity are required for full melanosome aggregation, but not dispersion; (3) partial aggregation that occurred despite myosin II or ROCK inhibition suggests a second component of aggregation that is dependent on cAMP signaling, but independent of ROCK and myosin II.     

A Late Jurassic plesiosaur in Antarctica: Evidence of the dispersion of marine fauna through the Trans-Erythraean Seaway?

During the Jurassic two main marine pathways might act as dispersion routes for vertebrates and invertebrates between Laurasia and Gondwana: the Caribbean Seaway (between North and South America) and the Trans-Erythraean Seaway (splitting Africa from India, Madagascar). The former has proven to be of relevance as a dispersion route for marine vertebrates and invertebrates between the Tethys and Pacific margin of Gondwana. Nevertheless, little is known about the role of the Trans-Erythraean Seaway as a vertebrate dispersion pathway. The Trans-Erythraean Seaway divides the eastern and western South of Gondwana landmasses in the so-called break-up of Gondwana and connects the Tethys Sea with the Palaeo-Pacific. We describe a newly recovered plesiosaur specimen from the Ameghino (= Nordensköld) Formation, Antarctic Peninsula, the first Jurassic plesiosaur from Antarctica. We discuss the importance of this record regarding the hypothesis of marine vertebrate dispersion through the Trans-Erythraean Seaway.     

Residence time distribution of diazepam in the isolated perfused rat liver. Analysis with the axial dispersion model.

The application of the axial dispersion model to diazepam hepatic elimination was evaluated using data obtained for impulse-response experiments with diazepam in the single-pass isolated perfused rat liver preparation. The transient form of the two-compartment dispersion model was applied to the output concentration versus time profile of diazepam after bolus input of a radiolabelled tracer into the hepatic portal vein (n = 4), providing DN and CLint estimates of 0.251 +/- 0.093 and 135 +/- 59 ml min-1, respectively. In contrast, the one-compartment form of the axial dispersion model, which assumes instantaneous transversal distribution of substance to the accessible spaces within the liver, could not adequately describe the residence time distribution (RTD) of diazepam. Furthermore, the magnitude of DN, a stochastic parameter which characterizes the axial spreading of solutes during transit through the liver, is similar to that determined for non-eliminated substances such as erythrocytes, albumin, sucrose and water. These findings suggest that the dispersion of diazepam in the perfused rat liver is determined primarily by the architecture of the hepatic microvasculature.     

Fluid particle diffusion through high-hematocrit blood flow within a capillary tube

Fluid particle diffusion through blood flow within a capillary tube is an important phenomenon to understand, especially for studies in mass transport in the microcirculation as well as in solving technical issues involved in mixing in biomedical microdevices. In this paper, the spreading of tracer particles through up to 20% hematocrit blood, flowing in a capillary tube, was studied using a confocal micro-PTV system. We tracked hundreds of particles in high-hematocrit blood and measured the radial dispersion coefficient. Results yielded significant enhancement of the particle diffusion, due to a micron-scale flow-field generated by red blood cell motions. By increasing the flow rate, the particle dispersion increased almost linearly under constant hematocrit levels. The particle dispersion also showed near linear dependency on hematocrit up to 20%. A scaling analysis of the results, on the assumption that the tracer trajectories were unbiased random walks, was shown to capture the main features of the results. The dispersion of tracer particles was about 0.7 times that of RBCs. These findings provide good insight into transport phenomena in the microcirculation and in biomedical microdevices.     

木质纤维素的定量测定及降解规律的初步研究

为了准确地测定稻草及其发酵物中纤维素、半纤维素、木质素的含量,通过差重法进行定量测定,并以此评价白腐菌株Pleurotus sapidus对稻草秸秆的降解状况,结果表明：利用差重法测定稻草发酵物中纤维素、半纤维索、木质素的百分含量是可行的,并能很好地评价白腐菌对稻草的降解规律,即降解过程中纤维素、半纤维素、本质素在前20d降解的很快,之后降解减缓,在50d内,纤维素被降解34．02％,半纤维素被降解56．29％,木质素被降解61．65％。     

Dielectric behaviour of frozen DNA in solution

In this article, measurements are reported on ice and frozen DNA solutions between 100 Hz–10 MHz. Pure ice is shown to exhibit single relaxation behaviour, which confirms previous work taken over a more restricted frequency range. The frozen DNA solution displays double‐dispersion behaviour. One dispersion centred around 3 kHz is due to a defect mechanism while the other, centred around 2 MHz, may be attributed to counterion flow through the water immediately adjacent to the DNA molecule. Bioelectromagnetics 20:40–45, 1999. © 1999 Wiley‐Liss, Inc.     

Influence of reactor geometry on mixing characteristics in a down flow jet loop bioreactor: newtonian and non-newtonian fluids

Mixing characteristics in the downcomer and the riser of a continuous down-flow jet loop bioreactor was studied with Newtonian and non-Newtonian fluids. The mixing parameters were determined through the curve fitting of the experimental impulse response data with the solution of one dimensional axial dispersion model. It was found that circulation number and axial dispersion coefficient increased with an increase in liquid flow rate and draft tube to column diameter ratio and the axial dispersion coefficient was comparatively higher in the riser. The circulation number increased with decrease in nozzle diameter. The model predicted the experimental data well within 8% deviation for both the systems (water and CMC). Correlations were obtained to predict axial dispersion coefficients in the riser and downcomer of the reactor.     

The protein kinase A-anchoring protein moesin is bound to pigment granules in melanophores

Major signaling cascades have been shown to play a role in the regulation of intracellular transport of organelles. In Xenopus melanophores, aggregation and dispersion of pigment granules are regulated by the second messenger cyclic AMP through the protein kinase A (PKA) signaling pathway. PKA is bound to pigment granules where it forms complexes with molecular motors involved in pigment transport. Association of PKA with pigment granules occurs through binding to A-kinase-anchoring proteins (AKAPs), whose identity remains largely unknown. In this study, we used mass spectrometry to examine an 80 kDa AKAP detected in preparations of purified pigment granules. We found that tryptic digests of granule protein fractions enriched in the 80 kDa AKAP contained peptides that corresponded to the actin-binding protein moesin, which has been shown to function as an AKAP in mammalian cells. We also found that recombinant Xenopus moesin interacted with PKA in vitro , copurified with pigment granules and bound to pigment granules in cells. Overexpression in melanophores of a mutant moesin lacking conserved PKA-binding domain did not affect aggregation of pigment granules but partially inhibited their dispersion. We conclude that Xenopus moesin is an AKAP whose PKA-scaffolding activity plays a role in the regulation of pigment dispersion in Xenopus melanophores.     

Beyond mean modelling: Bias due to misspecification of dispersion in Poisson‐inverse Gaussian regression

In clinical trials one traditionally models the effect of treatment on the mean response. The underlying assumption is that treatment affects the response distribution through a mean location shift on a suitable scale, with other aspects of the distribution (shape/dispersion/variance) remaining the same. This work is motivated by a trial in Parkinson's disease patients in which one of the endpoints is the number of falls during a 10‐week period. Inspection of the data reveals that the Poisson‐inverse Gaussian (PiG) distribution is appropriate, and that the experimental treatment reduces not only the mean, but also the variability, substantially. The conventional analysis assumes a treatment effect on the mean, either adjusted or unadjusted for covariates, and a constant dispersion parameter. On our data, this analysis yields a non‐significant treatment effect. However, if we model a treatment effect on both mean and dispersion parameters, both effects are highly significant. A simulation study shows that if a treatment effect exists on the dispersion and is ignored in the modelling, estimation of the treatment effect on the mean can be severely biased. We show further that if we use an orthogonal parametrization of the PiG distribution, estimates of the mean model are robust to misspecification of the dispersion model. We also discuss inferential aspects that are more difficult than anticipated in this setting. These findings have implications in the planning of statistical analyses for count data in clinical trials.     

Molecular dynamics simulation of dispersion improvement of graphene sheets in nanofluids by steric hindrance resulting from functional groups

Nanofluids are candidate materials for thermal management of heat transfer equipment. Practical applications of thermally enhanced nanofluids contribute to the reduction of weight of systems, leading to improved energy efficiency. Microsize particles sink into the systems because of gravity, therefore rendering the addition meaningless in terms of improving thermal properties. However, nanoparticles can be buoyant, leading to Brownian motion in the fluid, when they do not aggregate with each other. The most important factor in nanofluids is long-term stability of the dispersion in the fluid. Numerous studies have reported the dispersion stability; functional groups attached to nanoparticles play a role in causing steric hindrance and have an affinity for the surrounding fluid, resulting in preserving the dispersion. We investigate the structural effects on dispersion by molecular dynamics simulations of nanofluid containing graphene sheets with functional groups of varying lengths at the surface. The results demonstrate that short functional groups were too short to cause significant steric hindrance, while relatively longer functional groups tended to stack onto the graphene sheets, leading to trapping due to strong van der Waals interactions. Additionally, we discuss the minimum number of functional groups necessary for maintaining dispersion through calculations of the area of a single functional group.     

The cytoplast: A unit structure in chromatophores

We followed the translocation of identifiable pigment granules in living erythrophores through normal aggregation and dispersion and observed that they always return in dispersion to the same location relative to the whole pigment complex. This is interpreted to mean that each granule occupies a fixed position within a unit structure, the cytoplast. This position is retained even though the cytoplast undergoes dramatic reversals in form from ellipsoid to spheroid and back again with each aggregation and dispersion. The major structural components of the cytoplast, besides pigment granules, are microtubules and microtrabeculae. The latter constitute an irregular lattice that is confluent with microtubules and contains the pigment granules. In aggregation, the microtrabeculae shorten and seemingly contribute to the contraction of the entire cytoplast plus pigment. In dispersion, the microtrabeculae elongate in an apparent restructuring of the ellipsoidal cytoplast. The microtubules, however, persist in the cell cortex and appear to give radial direction to the pigment motion.     

Pulmonary arterial transit times

To begin to characterize the pulmonary arterial transport function we rapidly injected a bolus containing a radiopaque dye and a fluorescence dye into the right atrium of anesthetized dogs. The concentrations of the dye indicators were measured in the main pulmonary artery (fluoroscopically) and in a subpleural pulmonary arteriole (by fluorescence microscopy). The resulting concentration vs. time curves were subjected to numerical deconvolution and moment analysis to determine how the bolus was dispersed as it traveled through the arteriole stream tube from the main pulmonary artery to the arteriole. The mean transit time and standard deviation of the transport function from the main pulmonary artery to the arterioles studied averaged 1.94 and 1.23 s, respectively, and the relative dispersion (ratio of standard deviation to mean transit time) was approximately 64%. This relative dispersion is at least as large as those reported for the whole dog lung, indicating that relative to their respective mean transit times the dispersion upstream from the arterioles is comparable to that taking place in capillaries and/or veins. The standard deviations of the transport functions were proportional to their mean transit times. Thus the relative dispersion from the main pulmonary artery to the various arterioles studied was fairly consistent. However, there were variations in mean transit time even between closely adjacent arterioles, suggesting that variations in mean transit times between arteriole stream tubes also contribute to the dispersion in the pulmonary arterial tree.