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The Morphology and Equilibration of Levitated Secondary Organic Particles Under Controlled ConditionsGorkowski, Kyle J. 01 September 2017 (has links)
I advanced the understanding of particle morphology and its implications for the behavior and effects of atmospheric aerosol particles. I have developed new experimental methods for the Aerosol Optical Tweezers (AOT) system and expanded the AOT’s application into studying realistic secondary organic aerosol (SOA) particle phases. The AOT is a highly accurate system developed to study individual particles in real-time for prolonged periods of time. While previous AOT studies have focused on binary or ternary chemical systems, I have investigated complex SOA, and how they interact with other chemical phases, and the surrounding gas-phase. This work has led to new insights into liquid-liquid phase separation and the resulting particle morphology, the surface tension, solubility, and volatility of SOA, and diffusion coefficients of SOA phases. I designed a new aerosol optical tweezers chamber for delivering a uniformly mixed aerosol flow to the trapped droplet’s position. I used this chamber to determine the phase-separation morphology and resulting properties of complex mixed droplets. A series of experiments using simple compounds are presented to establish my ability to use the cavity enhanced Raman spectra to distinguish between homogenous single-phase, and phase-separated core-shell or partially-engulfed morphologies. I have developed a new algorithm for the analysis of whispering gallery modes (WGMs) present in the cavity enhanced Raman spectra retrieved from droplets trapped in the AOT. My algorithm improves the computational scaling when analyzing core-shell droplets (i.e. phase-separated or biphasic droplets) in the AOT, making it computationally practical to analyze spectra collected over many hours at a few Hz. I then demonstrate for the first time the capture and analysis of SOA on a droplet suspended in an AOT. I examined three initial chemical systems of aqueous NaCl, aqueous glycerol, and squalane at ~ 75% relative humidity. For each system I added α-pinene SOA – generated directly in the AOT chamber – to the trapped droplet. The resulting morphology was always observed to be a core of the initial droplet surrounded by a shell of the added SOA. By combining my AOT observations of particle morphology with results from SOA smog chamber experiments, I conclude that the α-pinene SOA shell creates no major diffusion limitations for water, glycerol, and squalane under humid conditions. My AOT experiments highlight the prominence of phase-separated core-shell morphologies for secondary organic aerosols interacting with a range of other chemical phases. The unique analytical capabilities of the aerosol optical tweezers provide a new approach for advancing the understanding of the chemical and physical evolution of complex atmospheric particulate matter, and the important environmental impacts of aerosols on atmospheric chemistry, air quality, human health, and climate change.
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CaMKII activation triggers persistent formation and segregation of postsynaptic liquid phase / CaMKIIの活性化によるシナプス後部液相の持続的な形成と分離Liu, Pin-Wu 23 March 2021 (has links)
京都大学 / 新制・課程博士 / 博士(医科学) / 甲第23115号 / 医科博第126号 / 新制||医科||8(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 伊佐 正, 教授 髙橋 良輔, 教授 井上 治久 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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Pathological Aggregation and Liquid-Liquid Phase Separation of TDP-43 in Neurodegenerative DiseaseBabinchak, William Michael 29 May 2020 (has links)
No description available.
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Investigation on liquid liquid phase separation of lysozyme by dynamic light scatteringPoggemann, Hanna-Friederike January 2021 (has links)
The liquid-liquid phase separation (LLPS) of biomolecules is a phenomenon which received a lot of attention in the last years because it is not only related to theformation of membraneless organelles but also to neurodegenerative diseases. Lysozyme is a globular protein that undergoes LLPS in a buffer salt system andfor that it is well investigated with several techniques like microscopy, dynamic lightscattering (DLS) or small-angle X-ray scattering. In this work we investigate the effect of temperature, solvent and sample con-centration on the diffusion coefficient, the hydrodynamic radius and the viscosity oflysozyme using a DLS setup. Furthermore, the influence of these parameters on thecluster formation is addressed. Finally, we investigate the question if the LLPS oflysozyme in a buffer environment effects the formation of dynamic clusters.
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Liquid-Liquid Phase Separation as a Modulator of Pathological Aggregation of TauBoyko, Solomiia 26 May 2023 (has links)
No description available.
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Investigation on Liquid-Liquid Phase Separation in Immunoglobulin G SolutionsJansson, Lovisa January 2023 (has links)
Liquid-liquid phase separation (LLPS) is an important phenomenon in soft condensed matter that explains many properties of membraneless organelles in living cells. The research on this topic is, therefore, a field with a wide range of applications such as biopharmacy and biomaterials. In this project, we investigate the LLPS of the antibody protein Immunoglobulin G (IgG) by analyzing the liquid dynamics of IgG solutions at a wide range of temperatures with dynamic light scattering (DLS). It was found that the slow component of the autocorrelation function increases with decreasing temperature below 0 °C. This can be attributed to either the number of protein clusters increasing as the sample approaches phase separation or LLPS droplets forming in the solution. LLPS was detected through optical microscopy, visualising the droplet formation in the IgG solution. This work confirms that LLPS can be detected for bovine IgG solutions without the presence of cosolvents and without water freezing in the sample.
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Crystal and Particle Engineering: Pharmaceutical Cocrystals through Antisolvent and Liquid-Liquid Phase Separation TechnologiesSajid, Muhammad A. January 2019 (has links)
The effects of polymer concentration and solvents on cocrystal morphology of
low solubility drugs were investigated, both of which had an impact. The
melting temperatures also decreased with increasing polymer concentration.
Placing the binding agent, benzene, at different interfaces induced
morphological changes, such as formation of porous cocrystals.
Previously liquid-liquid phase separation (LLPS) has been reported as a
hindrance in the crystallisation process impeding further development. A
phase diagram was constructed, and different phases were categorised into 4
types. After separation of the highly concentrated amorphous Oil Phase II, it
was prone to gradual crystallisation. Crystallisation took place over 30-60
minutes; this allowed the in-situ monitoring.
A novel cocrystallisation technique was developed; from (LLPS). Cocrystals of
indomethacin with saccharin and nicotinamide were obtained by mixing Oil
Phase II with the coformers.
In-situ monitoring by spectroscopic had gradual changes in spectra;
characteristic peaks increased in height and area with the formation of crystals
until the reaction was complete. With crystal formation, the XRD spectra
gradually had a sharper baseline due to a decrease in the amorphous
indomethacin. The photoluminescence (PL) spectra displayed several peaks
coupling into one large hump together with increasing intensity as the sample
crystallised. There was a shift in the peak absorbance of the pure drug crystals
obtained from LLPS and the indomethacin:saccharin cocrystal obtained from
LLPS.
Amorphous stabilisation was achieved by mixing polymer (PVP) with Oil
Phase II. There were no changes to the XRD diffractogram as the sample did
not undergo crystallisation.
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Interactions et assemblages de prolamines du blé / Interactions and assemblies of wheat prolaminsPincemaille, Justine 22 November 2018 (has links)
Ce travail de thèse vise à apporter des connaissances structurales et fonctionnelles sur les protéines du gluten. Pour cela, nous utilisons les concepts et méthodes de la physique des polymères et de la matière molle. Plus précisément, nous optimisons un protocole d’extraction basé sur la séparation de phases liquide-liquide. Ce dernier permet d’obtenir des isolats de protéines à différents rapports massiques gluténines/gliadines que nous étudions ensuite dans un solvant eau/éthanol 50/50 (v/v). Les résultats, montrent que les protéines se comportent comme des chaînes de polymères en solvant θ, en régime dilué et semi-dilué avec des tailles caractéristiques définis par diffusion de rayons X et de neutrons aux petits angles. De plus, 2 tailles d’objets sont distinguées en régime dilué par diffusion dynamique de la lumière: d’une part des protéines monomériques de l’ordre d’une dizaine de nanomètres associées aux et -gliadines et à des polymères de gluténines de faibles masses molaires et d’autre part des assemblages polymériques de l’ordre de 100 nm, principalement composés de ω-gliadines et polymères de gluténines de haute masse molaire. Ces assemblages sont mis en avant par une combinaison de mesures réalisées par chromatographie d’exclusion de taille et par fractionnement par flux de forces asymétrique et permettent de rationaliser les diagrammes de phases de ces mélanges protéiques, en fonction de la température. L’étude de la dynamique de séparation de phases de ces mélanges protéiques, par diffusion de rayons X aux petits angles, montre que celle-ci est pilotée par un phénomène de décomposition spinodale. Cette décomposition peut être arrêtée lors de trempes en température profondes mais également observée à toutes les températures de trempe, pour les échantillons les plus riches en gluténines, formant un gel dès le régime monophasique, d’après leur étude par rhéologie / The aim of this thesis is to provide structural and functional knowledge on wheat gluten proteins. For that, we use the physical methods and the concept of soft matter. We optimize an extraction protocol based on a liquid-liquid phase separation. With this protocol, we obtain protein batches with different glutenin/gliadin mass ratios, which we then study in a 50/50 water/ethanol solvent (v/v). We show that proteins behave like polymer chains in θ solvent in dilute and semi-dilute regime, whose characteristic size are extracted by small angle X-ray and neutron scattering. Moreover, two sizes of objects are evidenced in dilute regime by dynamic light scattering: monomeric proteins with a size around 10 nm which can be associated to α/β, and γ-gliadins and polymeric glutenins with low molecular weight and polymeric assemblies with a size around 100 nm composed of ω-gliadins and glutenins polymers with high molecular weight. These assemblies are revealed by a combination of size exclusion chromatography and asymmetric flow field flow fractionation and allow one to rationalize the phase diagrams of the protein mixtures with temperature. The study of the dynamics of the phase separation of these protein mixtures by small angle X-ray scattering shows that the phase separation proceeds through a spinodal decomposition phenomenon. An arrested phase separation is observed for deep quenches but also at all temperature quenches for the most glutenin rich samples, which are gels in the monophasic regime, as confirmed by rheology.
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Liquid-Liquid Phase Separation in an Isorefractive Polethylene Blend Monitored by Crystallization Kinetics and Crystal-Decorated Phase MorphologiesWang, Shujun 17 December 2008 (has links)
No description available.
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Mechanistic Studies of Human Immune Disease Relevant Genes and CRISPR Genome Editing Using Stem CellsYuan, Baolei 11 1900 (has links)
Stem cells, with the ability to self-renew and differentiate into intended cell types, are a valuable tool for disease modeling and mechanistic study. CRISPR-Cas9 has been widely used for genome editing due to its high efficiency and convenience. However, CRISPR-Cas9 has large-deletion safety issues that dramatically restrict its applications. Wiskott-Aldrich syndrome (WAS) is an inborn immunological disorder caused by WASP deficiency. WASP functions in the nucleus, which may help to understand WAS pathology, are poorly defined. Pannexin 1 (PANX1) forms large plasma membrane pores to exchange intracellular small molecules with the extracellular environment and functions in inflammatory processes. The regulatory mechanisms of the PANX1 channel remain obscure. In this dissertation, I focused on mechanistic studies of CRISPR-Cas9 genome editing, and two immune disease relevant genes, WASP and PANX1 using stem cell-derived immune cells.
We first found that CRISPR-induced large deletions (LDs) are predominantly mediated by the MMEJ repair pathway through statistical studies. Further, we found POLQ and RPA play vital roles in CRISPR-induced LDs. Modulation of POLQ and RPA can decrease CRISPR-induced LDs and increase HDR efficiency. Using three isogenic WAS iPSC models generated via gene editing, we successfully recapitulated WAS phenotypes, and for the first time, revealed that WASP regulates RNA splicing via epigenetically controlling the transcription of splicing factors and directly participating in the splicing machinery through a liquid-liquid phase separation process. We established a full-length human PANX1 (hPANX1) channel model via cryo-electron microscopy experiments and molecular dynamics simulation study, and found that hPANX1 channel is a homo-heptamer with both the N- and C-termini stretching deeply into the pore funnel. Functional studies of three selected residues support the new hPANX1 channel model and suggest the potential regulatory role of hPANX1 in pyroptosis upon immune responses.
Overall, the mechanistic studies of WASP, PANX1 and CRISPR genome editing revealed new roles of WASP in regulating RNA splicing, new functional insights of PANX1 in pyroptosis, and uncovered two critical players POLQ and RPA in CRISPR-induced LDs.
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