Potassium ion channels play a key role in the generation and propagation of action potentials. The S4-S5 linker peptide (L45) is believed to be responsible for the anesthetic/alcohol response of voltage-gated K+ channels. We investigated this region to define the structural basis of 1-alkanol binding site in dShaw2 K+ channel. L45 peptides derived from dShaw2 and hKv3.4 K+ channel, which, if part of the complete channel, demonstrate different sensitivity to 1-alcohols. Specifically, dShaw2 is alcohol sensitive and hKv3.4 is alcohol resistant. Structural analysis of L45 with NMR and CD suggested a direct correlation between alpha-helicity and the inhibition of dShaw2 channel by 1-butanol. We used CD and NMR to determine the structure of L45 peptides in micelles and vesicles. We measured spin-lattice relaxation time (T1) and determined the location and surface accessibility of L45 in micelles. These experiments confirm that L45 of dShaw2 adopts an α-helical conformation, partially buried in the membrane and parallel to the surface. The binding and accumulation of rev proteins to an internal loop of RRE (rev responsive element) of unspliced mRNA precursors is a key step of propagation of human immunodeficiency (HIV) virus. Molecules that interfere with this process can be expected to show anti-HIV activity. Our work is based on an assumption that zinc fingers could compete with rev proteins, therefore impeding the life cycle of HIV and stopping its infection. We studied the influence of different cations, anions, and the concentration of salts and osmolytes on the binding affinity with Polyacrylamide Gel Electrophoresis (PAGE) and Isothermal Titration Calorimetry (ITC). We conclude that the types of anions and/or cations and their concentrations affect the enthalpy and entropy of the binding interacitons. Using a gel assay, we confirm that there are three products in RNA-Protein reaction, and both EDTA and salts (and their concentrations) in the gel or samples interfere with RNA-protein complex mobility.

Potassium ion channels play a key role in the generation and propagation of action potentials. The S4-S5 linker peptide (L45) is believed to be responsible for the anesthetic/alcohol response of voltage-gated K+ channels. We investigated this region to define the structural basis of 1-alkanol binding site in dShaw2 K+ channel. L45 peptides derived from dShaw2 and hKv3.4 K+ channel, which, if part of the complete channel, demonstrate different sensitivity to 1-alcohols. Specifically, dShaw2 is alcohol sensitive and hKv3.4 is alcohol resistant. Structural analysis of L45 with NMR and CD suggested a direct correlation between alpha-helicity and the inhibition of dShaw2 channel by 1-butanol. We used CD and NMR to determine the structure of L45 peptides in micelles and vesicles. We measured spin-lattice relaxation time (T1) and determined the location and surface accessibility of L45 in micelles. These experiments confirm that L45 of dShaw2 adopts an α-helical conformation, partially buried in the membrane and parallel to the surface. The binding and accumulation of rev proteins to an internal loop of RRE (rev responsive element) of unspliced mRNA precursors is a key step of propagation of human immunodeficiency (HIV) virus. Molecules that interfere with this process can be expected to show anti-HIV activity. Our work is based on an assumption that zinc fingers could compete with rev proteins, therefore impeding the life cycle of HIV and stopping its infection. We studied the influence of different cations, anions, and the concentration of salts and osmolytes on the binding affinity with Polyacrylamide Gel Electrophoresis (PAGE) and Isothermal Titration Calorimetry (ITC). We conclude that the types of anions and/or cations and their concentrations affect the enthalpy and entropy of the binding interacitons. Using a gel assay, we confirm that there are three products in RNA-Protein reaction, and both EDTA and salts (and their concentrations) in the gel or samples interfere with RNA-protein complex mobility.

No description available.

No description available.

京都大学 / 新制・課程博士 / 博士(エネルギー科学) / 甲第24853号 / エネ博第462号 / 新制||エネ||87(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 野平, 俊之, 教授 萩原, 理加, 教授 佐川, 尚 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DFAM

<div>Potassium-ion batteries (PIBs) recently emerged as a next-generation energy storage technology, utilizing abundant and inexpensive potassium as the charge carrier cation. PIBs operate by an analogous mechanism to lithium-ion batteries (LIBs), with reversible potassium intercalation in anode and cathode through an inorganic salt - organic solvent electrolyte medium. Despite its larger size, potassium exhibits several electrochemical advantages over sodium, including a higher affinity for intercalation into graphitic (carbonaceous) anodes, forming a stage-one KC₈ structure in graphite for a specific capacity of 279 mAh g^-1. This thesis aims to provide a thorough foundation for PIB carbon anodes, through a comprehensive experimental approach combining electrode synthesis, advanced material characterization and electrochemical-analytical techniques.</div><div><br></div><div>Safety concerns have consistently plagued LIBs despite almost three decades of widespread commercialization. Thermal runaway of LIBs can initiate as early as 80°C from exothermic breakdown of the solid electrolyte interphase (SEI) layer that covers the carbon anode surface. The subsequent reaction of lithiated carbon with electrolyte solvent leads to cathode decomposition and oxygen release for cell gassing and combustion. This thesis investigates the thermal runaway behavior of graphite anode for PIBs via differential scanning calorimetry analysis, determining the effect of electrode material, state-of-charge, and cycling history on heat generation. Notably, the PIB system emits significantly less heat overall than for LIBs, albeit an earlier and more intense onset reaction at 100°C raises safety concerns. Strategies to mitigate this exothermic reaction are presented, including electrode binder manipulation to improve graphite particle coverage and enhance SEI layer stability.</div><div><br></div><div>To further evaluate the practicality of PIBs, the electrochemical behavior of graphite anode was investigated from 0 - 40°C operating temperature, in comparison to standard LIBs. The poor rate capability of potassium is attributed to sluggish solid-state diffusion and augmented cell impedance, where 3-electrode studies revealed dramatic polarization of the potassium metal counter electrode at low temperatures. Accelerated cell aging at elevated temperatures is attributed to SEI layer growth induced by the 61% volumetric expansion of graphite during potassiation, as well as the extreme reactivity of potassium metal. A full-cell system with a Prussian blue nanoparticle cathode and graphite anode showed enhanced rate performance at low temperatures by removing potassium metal counter electrode. These results provide valuable mechanistic insight for potassium intercalation in graphite and offer a practical evaluation of temperature dependent electrochemical performance for PIBs.</div><div><br></div><div>Supplementary research includes the exploration of carbon nanofibers electrospun from polyacrylonitrile precursor with subsequent pyrolysis as PIB anode. The design of an amorphous, low density carbon with a nanoscale one dimensional morphology enables mitigation of the 61% volumetric expansion of graphite during potassiation. Remarkable stability (2000 charge-discharge cycles) is thus achieved by preventing electrode pulverization, SEI layer growth, and impedance rise during cycling. Electrochemical analysis revealed a pseudo-capacitance mechanism, enabling rapid charging through surface storage of potassium that could be enhanced by surface functionalization via plasma oxidation treatment. Moreover, two dimensional MXene transition carbonitride sheets were explored as PIB anode with X-ray diffraction and X-ray photoelectron spectroscopy used to study structural changes during potassium insertion.</div><div><br></div><div>Finally, the effect of particle morphology was investigated for LIB carbon anodes, wherein commercial graphite is compared with synthesized spherical and spiky carbons. Intercalation dynamics, side reaction rates (e.g. SEI growth), self-heating, and thermal runaway behavior were studied through a combination of electrochemical analysis and modeling by a finite volume method. Spherical particles outperform irregular commercial graphite by eliminating unstructured inhomogeneities that lead to non-uniform current distributions. Interestingly, spiky particles offer a nontrivial response, where the ordered irregularities enhance intercalation dynamics to prevent degradation at extreme operating conditions. These findings emphasize the importance of tailoring particle morphology and structure in promoting desired LIB behavior and suppressing unwanted problems.</div>

京都大学 / 新制・課程博士 / 博士(エネルギー科学) / 甲第23295号 / エネ博第420号 / 新制||エネ||80(附属図書館) / 京都大学大学院エネルギー科学研究科エネルギー基礎科学専攻 / (主査)教授 萩原 理加, 教授 野平 俊之, 教授 坂口 浩司 / 学位規則第4条第1項該当 / Doctor of Energy Science / Kyoto University / DFAM

No description available.

In view of the increasing global challenge of bacterial resistance, there exists an urgent need for the rationale development of antibacterial compounds with either novel or multiple mechanisms of action. Two chalcone-derivatives, F1 and F23, demonstrated MICs within the range of 16 to >512 μg/ml against two plant pathogens (P. caratovoram and C. michiganensis subsp. michiganensis) as well as important clinical bacterial species. Both compounds displayed an MIC of 32 μg/ml against quinolone-resistant S. aureus. Whilst possessing weak activities individually, each semi-synthetic agent displayed notable synergistic action with polymyxin B against S. aureus, C. violaceum, E. coli and Ps. aeruginosa, thereby recording FICs within the range of <0.093 to 2 that indicated the existence of synergism in some instance. These chalcone compounds applied with polymyxin B displayed a notable FICindex of <0.093 against the Neisseriaceae C. violaceum, and a potential noteworthy capacity to extend the spectrum of activity of the latter antibiotic to include Gram-positive S. aureus species. F1 inhibited staphylococcal replication in broth and the combination of either of both chalcone-derivatives with polymyxin B instituted a metabolic blockage in S. aureus and other bacterial species as determined through a modified MTT reduction assay. The combined agents inflicted major disruptions to the S. aureus cytoplasmic membrane bilayer as evidenced by the release of intracellular potassium as well as the influx of Sytox Green fluorescent stain. Notable levels of cell membrane potential dissipation, leakage of intracellular potassium ions and blockage of reducing enzymes activities occurred within the first 30 minutes, well in advance of significant loss in cell viability that was recorded usually after 4 – 8 hours, suggesting these activities were prerequisites to cell death. In erythrocyte lysis assay, the synergistic combinations of 128 μg/ml of either of both chalcone derivatives with 128 μg/ml polymyxin B displayed the lowest degree of haemolysis, followed by that occurring with 32 μg/ml of the chalcone-derivatives combined with 256 μg/ml of the polypeptide antibiotic. In conclusion, further structure activity modifications aimed at improving the aqueous solubility of these chalcone-derivatives as well as the antibacterial activity recorded for certain combination concentrations of polymyxin B with either of these semi-synthetic agents may be required before considerations are made for the possibility for potential external formulations. Such preparations may include antiseptic creams, lotions, ointments, as well as aerosols that can be applied with nebulizers in targeted delivery for such cases like cystic fibrosis.

Cardiac arrhythmias are caused by a disruption of the normal initiation or propagation of electrical impulses in the heart. Hundreds of mutations in genes encoding ion channels or ion channel regulatory proteins are linked to congenital arrhythmia syndromes that increase the risk for sudden cardiac death. This dissertation focuses on how mutations in a gene (KCNQ1) that encodes a voltage-gated K+ ion channel (Kv7.1) can disrupt proper channel function and lead to abnormal repolarization of atrial and ventricular cardiomyocytes. In the heart, Kv7.1 coassembles with a regulatory protein to conduct the slowly activating delayed rectifier K+ current (IKs). Loss-of-function KCNQ1 mutations are linked to type 1 long QT syndrome (LQT1), and typically decrease IKs, which can lead to ventricular action potential (AP) prolongation. In patients, LQT1 is often characterized by an abnormally long corrected QT (QTc) interval on an electrocardiogram (ECG), and increases the risk for polymorphic ventricular tachycardias. KCNQ1 mutations are also linked to atrial fibrillation (AF), but cause a gain-of-function phenotype that increases IKs. Surprisingly, patients diagnosed with both LQT1 and AF are increasingly identified as genotype positive for a KCNQ1 mutation. The first aim of this dissertation was to determine a unique functional phenotype of KCNQ1 mutations linked to both arrhythmia syndromes by functional analyses via the whole-cell patch clamp technique in HEK293 cells. A proportion of patients with LQT1-linked KCNQ1 mutations do not have abnormal QTc prolongation known as latent LQT1. Interestingly, exercise can reveal abnormal QTc prolongation in these patients. During exercise, beta-adrenergic activation stimulates PKA to phosphorylate Kv7.1, causing an increase in IKs to prevent ventricular AP prolongation. Therefore, the second aim of this dissertation was to determine a molecular mechanism of latent LQT1 through functional analyses in HEK293 cells while incorporating pharmacological and phosphomimetic approaches to study PKA regulation of mutant Kv7.1 channels. The findings in this dissertation provide new insight into how KCNQ1 mutations disrupt the function of Kv7.1 in a basal condition or during beta-adrenergic activation. Also, this dissertation suggests these approaches will improve patient management by identifying mutation specific risk factors for patients with KCNQ1 mutations.