12 research outputs found

    7.胃管アレルギーに於ける腸粘膜スメアーの研究(第二報)(第415回千葉医学会例会,第63回日本小児科学会千葉地方会総会)

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    Extended representative data set of lysosomal positioning. (a) Schematic representation of the feature “MAX Contour Position” used to quantify lysosomal positioning. (b) Representative LAMP1 immunofluorescence images for different ranges of the feature “LAMP1 MAX Contour Position” in HeLa cells treated as in Fig. 6. (JPG 3911 kb

    In silico photoactivation to quantify transmission dynamics in mitochondrial networks.

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    <p>(A) Schematic representation of an <i>in silico</i> photoactivation. 10% of mitochondrial mass is assigned a GFP label, which transmit through the cell upon a fusion event: fusion of a mitochondrion with a GFP-labeled mitochondrion, results in a parent GFP-labeled mitochondrion, which upon fission results in two GFP-labeled daughter mitochondria. (B) Transmission dynamic of photoactivated GFP across the mitochondria population moving with free motion (ϑ<sub>1</sub> ϵ [0°, 360°]), starting from 10% of labeled mitochondria (green). Simulations were performed with total mitochondrial masses of 100 and 300 and three different fusion/fission probability rates: F<sub>p</sub>/f<sub>p</sub> = 20%/80% (dotted line), F<sub>p</sub>/f<sub>p</sub> = 50%/50% (solid line) and F<sub>p</sub>/f<sub>p</sub> = 80%/20% (dashed line). 100 simulations were performed each and plots represent the average values normalized to the total mass at time point 0. Blue regions indicate the time to 50% network transmission with fusion/fission probability rates of 50%/50%. (C) Same as (B) but with restricted movement, ϑ<sub>1</sub> ϵ [0°, 10°]. Pink regions indicate the time to 50% network transmission with fusion/fission probability rates of 50%/50%. (D) Effect of the mitochondria mobility on transmission dynamics in the case of free movement (blue circles, line) and restricted movement (pink triangles, line). Detailed representation of the time needed to the GFP labeled mitochondrial population to exceed the unlabeled population for an initial total mitochondrial mass of 300 with a fixed fusion/fission probability rate of 50%/50% and three different velocities: v<sub>l</sub> ϵ [0, 0.44] μms<sup>-1</sup>, v<sub>h</sub> ϵ [0, 1] μms<sup>-1</sup> and v, equal to v<sub>l</sub> in the perinuclear region and to v<sub>h</sub> outside.</p

    Sensitivity analysis of the transmission dynamics model.

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    <p>(A) Effect of parameters perturbation on transmission dynamic. Bar graph represents Partial Rank Correlation Coefficient values for each parameter-output response pairing resulting from a Latin-hypercube sampling of 500 parameter value sets (parameter ranges are described in section “Transmission dynamics of mitochondrial network connectivity”). For each parameter set the model was performed over 6 hours. (B) Results of the eFast method used to partition variance in simulation results between parameters. Solid Bars: sensitivity index (Si)–the fraction of output variance explained by the value assigned to that parameter when parameter of interest; Dashed Bars: total sensitivity index (STi)–the variance caused by higher-order non-linear effects between that parameter and the other explored (includes value of Si). Error bars are standard error over four resample curves. Parameter values sets were generated using the sinusoidal sampling approach. 65 parameters were obtained from each of the 4 resampling curves, producing 2080 parameter value sets (260 per parameter).</p

    Impact of fusion and fission probabilities and mitochondrial mass on mitochondrial subpopulations.

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    <p>(A) Time course response of mitochondrial subpopulations for low total mass (<i>M</i> = 100; 5% of the whole cell area). Three mitochondrial subpopulations are considered: small mass (orange line), medium mass (blue line) and large mass (green line). The partial mass reports the sum of the masses from all mitochondria in a given subpopulation divided by the total mitochondrial mass. Lines display the mean among 100 simulations, and shaded regions display the standard deviation. Small graphs underneath show the evolution of the first derivative of mitochondria with large mass (green line). Three different fusion/fission probability rates are used: 20%/80%, 50%/50% and 80%/20%. Pie charts report the partial mass (in terms of percentages) for the subpopulations at the final simulation time (2 hours). (B) Same as (A) but with a total mitochondrial mass of 200 (10% of the whole cell area). (C) Same as (A) but with a total mitochondrial mass of 300 (15% of the whole cell area).</p

    Interaction between mitochondrial health and cellular energetic state, based on the energetic stress.

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    <p>(A) Evolution of mitochondrial mass (grey lines) over time subjected to four different damage signals (black line). Line graphs display mean value and standard deviation (shaded regions) of 100 simulations for an initial total mitochondrial mass of 300 normalized to the total mass at time point 0. Parameter values obtained from the optimization were used, in particular, fusion probability = fission probability = 50%, fusion frequency = fission frequency = 5 minutes, biogenesis probability = 22.1%, biogenesis frequency = 28.9 minutes, receptor threshold = 12 minutes, damage threshold = 12.4 minutes, degradation frequency = 5.7 minutes and damage probabilities: 0%, 10%, 30% and 50%. Simulations were performed for a total of 12 hours. Blue region indicates the area in which biogenesis is dominant while red region indicates the area in which degradation is dominant. (B) Schematic representation of the link between the number of healthy and damaged mitochondria with the energetic stress (E<sub>S</sub>) of the cell. (C) Schematic representation of the connection between energetic (red line) stress and fission (dark green line) and biogenesis (orange line) probabilities. (D) Line graphs display mean value and standard deviation (shaded regions) of 100 simulations of probabilities of fusion (dark green line), fission (light green line), biogenesis (orange line) and damage signal (black line). Initial values assigned to the model: fusion probability = fission probability = 50%, fusion frequency = fission frequency = 5 minutes, biogenesis probability = 22.1%, biogenesis frequency = 28.9 minutes, receptor threshold = 12 minutes, damage threshold = 12.4 minutes, degradation frequency = 5.7 minutes. All the simulations were performed for a total time of 24 hours. (E) Line graphs display mean value and standard deviation (shaded area) of 100 simulations for an initial mitochondrial mass of 300 of the total mitochondrial mass (grey line) and three mitochondria subpopulations: small mass (orange line), medium mass (blue line) and big mass (green line) subjected to five different damage signals (10% and 20%) every two hours for one hour. The plot represents the evolution of the total mass and of three mitochondrial subpopulations normalized to the total mass at time point 0. Initial values assigned to the model: fusion probability = fission probability = 50%, fusion frequency = fission frequency = 5 minutes, biogenesis probability = 22.1%, biogenesis frequency = 28.9 minutes, receptor threshold = 12 minutes, damage threshold = 12.4 minutes, degradation frequency = 5.7 minutes. All the simulations were performed for a total time of 24 hours. (F) Line graphs display mean value and standard deviation (shaded regions) of 100 simulations of bioenergetics stress (red line). Initial values assigned to the model: fusion probability = fission probability = 50%, fusion frequency = fission frequency = 5 minutes, biogenesis probability = 22.1%, biogenesis frequency = 28.9 minutes, receptor threshold = 12 minutes, damage threshold = 12.4 minutes, degradation frequency = 5.7 minutes. All the simulations were performed for a total time of 24 hours. (G) Same as (D) but with mitochondria subjected to five different damage signals (40%) every two hours for one hour. (H) Same as (E) but with mitochondria subjected to five different damage signals (40%) every two hours for one hour. (I) Same as (F) but with mitochondria subjected to five different damage signals (40%) every two hours for one hour.</p

    Integration of mitochondrial biogenesis through parameter fitting.

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    <p>(A) Schematic representation of mitochondrial fusion, fission and biogenesis events. Biogenesis is represented by frequency (B<sub>f</sub>) and probability (B<sub>p</sub>) parameters, and can occur in all mitochondria with a mass less than M<sub>max</sub> = 3.0 μm<sup>2</sup>. (B) Basic flow chart of the model representing fusion, fission and biogenesis events. Shaded boxes indicate the main processes of the model (fusion, fission and biogenesis) with governing equation of the biogenesis process. Time step used in all the simulation, Δt = 1 sec. (C) Maximization of the fitness function O<sub>B</sub> within 100 iterations, reaching a value of -0.049, using a classic genetic algorithm (settings described in the Methods section). (D) Mean of 100 simulations of mitochondrial mass growth over 48 hours, beginning with a mitochondrial mass of 1 and using the parameter values obtained from the optimization (C), biogenesis probability = 22.1% and biogenesis frequency = 28.9 minutes.</p
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