GABA release by hippocampal astrocytes

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Parametric Analysis for Monitoring 2D Kinetics of Receptor-Ligand binding

Thermal fluctuation approach is widely used to monitor association kinetics of surface-bound receptor-ligand interactions. Various protocols such as sliding standard deviation (SD) analysis (SSA) and Page's test analysis (PTA) have been used to estimate two-dimensional (2D) kinetic rates from the time course of displacement of molecular carrier. In the current work, we compared the estimations from both SSA and modified PTA using measured data from an optical trap assay and simulated data from a random number generator. Our results indicated that both SSA and PTA were reliable in estimating 2D kinetic rates. Parametric analysis also demonstrated that such the estimations were sensitive to parameters such as sampling rate, sliding window size, and threshold. These results furthered the understandings in quantifying the biophysics of receptor-ligand interactions.

IL-8-induced L-selectin shedding regulates its binding kinetics to PSGL-1

L-selectin plays a crucial role in inflammation cascade by initiating the tethering and rolling of leukocytes on endothelium wall. While many L-selectin molecules are rapidly shed from the cell surface upon activation, the remaining membrane-anchored L-selectin may still play an important role in regulating leukocyte rolling and adhesion with different binding kinetics. Here we developed an in vitro model to activate Jurkat cells via interlukin-8 (IL-8) and quantified the two-dimensional (2D) binding kinetics, using a micropipette aspiration assay, of membrane-anchored L-selectin to P-selectin glycoprotein ligand 1 (PSGL-1) ligand coupled onto human red blood cells (RBCs). The data indicated that L-selectin shedding reduced the amount of membrane-anchored L-selectin and lowered both its reverse and forward rates. These results suggested that the rolling dynamics of activated leukocytes was determined by two opposite impacts: reducing the surface presentation would enhance the rolling but lowering the kinetic rates would decrease the rolling. This finding provides a new insight into understanding how L-selectin shedding regulates leukocyte rolling and adhesion.

Numerical Study On Transient Flow In The Deep Naturally Fractured Reservoir With High Pressure

According to the experimental results and the characteristics of the pressure-sensitive fractured formation, a transient flow model is developed for the deep naturally-fractured reservoirs with different outer boundary conditions. The finite element equations for the model are derived. After generating the unstructured grids in the solution regions, the finite element method is used to calculate the pressure type curves for the pressure-sensitive fractured reservoir with different outer boundaries, such as the infinite boundary, circle boundary and combined linear boundaries, and the characteristics of the type curves are comparatively analyzed. The effects on the pressure curves caused by pressure sensitivity module and the effective radius combined parameter are determined, and the method for calculating the pressure-sensitive reservoir parameters is introduced. By analyzing the real field case in the high temperature and pressure reservoir, the perfect results show that the transient flow model for the pressure-sensitive fractured reservoir in this paper is correct.

Investigation of transient process and steady output of lasing without inversion

The transient evolution processes and steady outputs of continuous wave lasing without inversion (LWI) and self-pulsing LWI in a resonant open V type three-level system are studied. It was found that the two kinds of LWI have some obvious differences not only from the steady outputs but also from the transient evolution processes. The effects of the unsaturated gain coefficient, cavity loss coefficient, ratio of the atomic injection rates and atomic exit rate on the transient evolution processes and steady outputs are discussed.

Finding needles in the haystack : a search for AM CVn systems using the Palomar Transient Factory

The AM CVn systems are a rare class of ultra-compact astrophysical binaries. With orbital periods of under an hour and as short as five minutes, they are among the closest known binary star systems and their evolution has direct relevance to the type Ia supernova rate and the white dwarf binary population. However, their faint and rare nature has made population studies of these systems difficult and several studies have found conflicting results.

I undertook a survey for AM CVn systems using the Palomar Transient Factory (PTF) astrophysical synoptic survey by exploiting the "outbursts" these systems undergo. Such events result in an increase in luminosity by a factor of up to two-hundred and are detectable in time-domain photometric data of AM CVn systems. My search resulted in the discovery of eight new systems, over 20% of the current known population. More importantly, this search was done in a systematic fashion, which allows for a population study properly accounting for biases.

Apart from the discovery of new systems, I used the time-domain data from the PTF and other synoptic surveys to better understand the long-term behavior of these systems. This analysis of the photometric behavior of the majority of known AM CVn systems has shown changes in their behavior at longer time scales than have previously been observed. This has allowed me to find relationships between the outburst properties of an individual system and its orbital period.

Even more importantly, the systematically selected sample together with these properties have allowed me to conduct a population study of the AM CVn systems. I have shown that the latest published estimates of the AM CVn system population, a factor of fifty below theoretical estimates, are consistent with the sample of systems presented here. This is particularly noteworthy since my population study is most sensitive to a different orbital period regime than earlier surveys. This confirmation of the population density will allow the AM CVn systems population to be used in the study of other areas of astrophysics.

Ultrafast transient ring-shaped population grating

We demonstrate an ultrafast transient, ring-shaped population grating induced by an ultrashort hollow Gaussian laser bullet by solving the three-dimensional full-wave Maxwell-Bloch equations. Through adjusting the beam waist and the area of the pulse, we can control the number of lines and the period of the grating. Based on this coherent control scheme, a door to produce gratings with complex transverse structure is opened.

Guiding and confining fast electrons by transient electric and magnetic fields with a plasma inverse cone

The fast electron propagation in an inverse cone target is investigated computationally and experimentally. Two-dimensional particle-in-cell simulation shows that fast electrons with substantial numbers are generated at the outer tip of an inverse cone target irradiated by a short intense laser pulse. These electrons are guided and confined to propagate along the inverse cone wall, forming a large surface current. The propagation induces strong transient electric and magnetic fields which guide and confine the surface electron current. The experiment qualitatively verifies the guiding and confinement of the strong electron current in the wall surface. The large surface current and induced strong fields are of importance for fast ignition related researches.

Electron flow through cytochrome P450

The cytochromes P450 (P450s) are a remarkable class of heme enzymes that catalyze the metabolism of xenobiotics and the biosynthesis of signaling molecules. Controlled electron flow into the thiolate-ligated heme active site allows P450s to activate molecular oxygen and hydroxylate aliphatic C–H bonds via the formation of high-valent metal-oxo intermediates (compounds I and II). Due to the reactive nature and short lifetimes of these intermediates, many of the fundamental steps in catalysis have not been observed directly. The Gray group and others have developed photochemical methods, known as “flash-quench,” for triggering electron transfer (ET) and generating redox intermediates in proteins in the absence of native ET partners. Photo-triggering affords a high degree of temporal precision for the gating of an ET event; the initial ET and subsequent reactions can be monitored on the nanosecond-to-second timescale using transient absorption (TA) spectroscopies. Chapter 1 catalogues critical aspects of P450 structure and mechanism, including the native pathway for formation of compound I, and outlines the development of photochemical processes that can be used to artificially trigger ET in proteins. Chapters 2 and 3 describe the development of these photochemical methods to establish electronic communication between a photosensitizer and the buried P450 heme. Chapter 2 describes the design and characterization of a Ru-P450-BM3 conjugate containing a ruthenium photosensitizer covalently tethered to the P450 surface, and nanosecond-to-second kinetics of the photo-triggered ET event are presented. By analyzing data at multiple wavelengths, we have identified the formation of multiple ET intermediates, including the catalytically relevant compound II; this intermediate is generated by oxidation of a bound water molecule in the ferric resting state enzyme. The work in Chapter 3 probes the role of a tryptophan residue situated between the photosensitizer and heme in the aforementioned Ru-P450 BM3 conjugate. Replacement of this tryptophan with histidine does not perturb the P450 structure, yet it completely eliminates the ET reactivity described in Chapter 2. The presence of an analogous tryptophan in Ru-P450 CYP119 conjugates also is necessary for observing oxidative ET, but the yield of heme oxidation is lower. Chapter 4 offers a basic description of the theoretical underpinnings required to analyze ET. Single-step ET theory is first presented, followed by extensions to multistep ET: electron “hopping.” The generation of “hopping maps” and use of a hopping map program to analyze the rate advantage of hopping over single-step ET is described, beginning with an established rhenium-tryptophan-azurin hopping system. This ET analysis is then applied to the Ru-tryptophan-P450 systems described in Chapter 2; this strongly supports the presence of hopping in Ru-P450 conjugates. Chapter 5 explores the implementation of flash-quench and other phototriggered methods to examine the native reductive ET and gas binding events that activate molecular oxygen. In particular, TA kinetics that demonstrate heme reduction on the microsecond timescale for four Ru-P450 conjugates are presented. In addition, we implement laser flash-photolysis of P450 ferrous–CO to study the rates of CO rebinding in the thermophilic P450 CYP119 at variable temperature. Chapter 6 describes the development and implementation of air-sensitive potentiometric redox titrations to determine the solution reduction potentials of a series of P450 BM3 mutants, which were designed for non-native cyclopropanation of styrene in vivo. An important conclusion from this work is that substitution of the axial cysteine for serine shifts the wild type reduction potential positive by 130 mV, facilitating reduction by biological redox cofactors in the presence of poorly-bound substrates. While this mutation abolishes oxygenation activity, these mutants are capable of catalyzing the cyclopropanation of styrene, even within the confines of an E. coli cell. Four appendices are also provided, including photochemical heme oxidation in ruthenium-modified nitric oxide synthase (Appendix A), general protocols (Appendix B), Chapter-specific notes (Appendix C) and Matlab scripts used for data analysis (Appendix D).

Transient population and polarization gratings induced by (1+1)-dimensional ultrashort dipole soliton

An ultrafast transient population grating induced by a (1+1)-dimensional, ultrashort dipole soliton is demonstrated by solving the full-wave Maxwell-Bloch equations. The number of lines and the period of the grating can be controlled by the beam waist and the area of the pulse. Of interest is that a polarization grating is produced. A coherent control scheme based on these phenomena can be contemplated as ultrafast transient grating techniques.

Studies in nuclear reactor dynamics : I. The accuracy of point kinetics. II. The effect of delayed neutrons on the spectrum of the group-diffusion operator

This thesis is a theoretical work on the space-time dynamic behavior of a nuclear reactor without feedback. Diffusion theory with G-energy groups is used.

In the first part the accuracy of the point kinetics (lumped-parameter description) model is examined. The fundamental approximation of this model is the splitting of the neutron density into a product of a known function of space and an unknown function of time; then the properties of the system can be averaged in space through the use of appropriate weighting functions; as a result a set of ordinary differential equations is obtained for the description of time behavior. It is clear that changes of the shape of the neutron-density distribution due to space-dependent perturbations are neglected. This results to an error in the eigenvalues and it is to this error that bounds are derived. This is done by using the method of weighted residuals to reduce the original eigenvalue problem to that of a real asymmetric matrix. Then Gershgorin-type theorems .are used to find discs in the complex plane in which the eigenvalues are contained. The radii of the discs depend on the perturbation in a simple manner.

In the second part the effect of delayed neutrons on the eigenvalues of the group-diffusion operator is examined. The delayed neutrons cause a shifting of the prompt-neutron eigenvalue s and the appearance of the delayed eigenvalues. Using a simple perturbation method this shifting is calculated and the delayed eigenvalues are predicted with good accuracy.