Effects of mutations at position 313 of Kir3.2 on basal activity.

<div><p>Comparison of the ratios of the basal current of Kir3.2 WT and mutant channels.</p> <p>The current amplitudes of Kir3.2 WT and mutant channels in the absence of ACh (I_basal) were divided by those in the presence of 10 μM ACh (I_total) in every oocyte. The I_basal/I_total ratio was plotted against the amplitude of I_total for the WT and each mutant. The plot shows a linear relationship when the I_total is less than 10 μA.</p></div

Crystal structure of the Kir3.2 E236R mutant.

<div><p><i>A</i>. Comparison of the crystal structures of the cytoplasmic domains (CPDs) of WT and E236R channels. The CPD of the E236R mutant (blue) behaves as a tetramer and has a structure similar to that of the WT in a closed conformation (gray). The βC and βD strands (CD loop; yellow) and HI loop (orange) are denoted by circles in 1 subunit and traced with lines in the next subunit.</p> <p><i>B</i>. Enlarged view of the subunit interface at position 236. An arginine introduced at site 236 of one subunit (blue) forms hydrogen bonds with the carbonyl oxygen atoms of Gly312 on the βH strand and Cys321 on the βI strand of the adjacent subunit (gray). The residues crucial for the interaction are shown as sticks.</p> <p><i>C</i>. The β-bulge in the βD strand. The β sheet observed from the pore is shown with ribbons and sticks. The side chains of Glu236 and Met313 are shown with dots. The β-type hydrogen bonds are disconnected between Glu236 and Gly237 on the βD strand and Gly312 on βH strand. The dashed line indicates the hydrogen bond between the main chain atoms.</p></div

Inside-out patch recordings of Kir3.2 WT and mutant channels.

<p>Inside-out membrane patches were obtained from HEK293T cells expressing either Kir3.2 WT (<i>A</i>), M313G (<i>B</i>) or M313T mutant (<i>C</i>) channels. Experiments were conducted in symmetric 135 mM K⁺ solutions with a holding potential of -100 mV. Perfusion of ATP was for the generation of PIP₂ at the inner leaflet of the excised patch membranes. The protocol used for perfusion of substances to the intracellular side of the patch membrane is indicated by bars above the current traces. </p

Effects of mutations at Glu236 and Met313 on m₂R-dependent activation.

<div><p><i>A</i>. Typical current traces of Kir3.2 wild-type (WT) and mutant channels recorded in the presence of various concentrations of ACh. The current responses were elicited by a test pulse at −120 mV for 0.6 s followed by a voltage step to +40 mV for 0.6 s at an interval of 0.1 s at −20 mV. This sequence was repeated every 3 s. The current in the presence of 3 mM Ba²⁺ was subtracted from each trace. The arrowheads indicate the zero current levels.</p> <p><i>B</i>, <i>C</i>. Current response of Kir3.2 WT and mutant channels. ACh-induced K⁺ currents recorded at the end of the test pulse to −120 mV were normalized to that measured in the presence of 10 μM ACh. Error bars indicate the standard error of the mean. The current responses of the WT and mutants were fit using the Hill equation as described in the Materials and Methods section.</p></div

Effects of mutations at position 313 of Kir3.2 on single-channel currents.

<div><p><i>A</i>. Single-channel recording of the indicated channel at −100 mV. The patch recording in the cell-attached configuration was obtained from HEK293T cells expressing Kir3.2 WT, M313G, and M313T. The holding potential was −100 mV. Open dwell time histograms for each channel are shown at the right side of the trace and are fit with a single exponential function. The single-channel current amplitudes of the WT and M313G mutant are plotted against the membrane potentials.</p> <p><i>B</i>. Effects of substitutions of threonine 301 in Kir2.1.</p> <p>Met301 in Kir2.1 is equivalent to Met313 in Kir3.2. The substitution of threonine at Met301 in Kir2.1 resulted in a spiky opening with variable conductance. Arrowheads indicate the zero current level.</p></div

Emission Tuning of Heteroleptic Arylborane–Ruthenium(II) Complexes by Ancillary Ligands: Observation of Strickler–Berg-Type Relation

Novel heteroleptic arylborane–ruthenium­(II) complexes having a series of ancillary ligands L′ ([Ru­(B₂bpy)­L′₂]²⁺) in CH₃CN showed low-energy/intense metal-to-ligand charge transfer (MLCT)-type absorption and intense/long-lived emission compared to the reference complexes. The spectroscopic and photophysical properties of [Ru­(B₂bpy)­L′₂]²⁺ were shown to be manipulated synthetically by the electron-donating ability of the ancillary ligand(s). The intense and long-lived emission observed for [Ru­(B₂bpy)­L′₂]²⁺ in CH₃CN at 298 K is responsible for the accelerated radiative and decelerated nonradiative decay processes, which are controllable through the electronic structures of the ancillary ligand(s). On the basis of the present systematic study, furthermore, we succeeded in demonstrating the Strickler–Berg-type relation between the molar absorption coefficients of the MLCT bands and the radiative rate constants of the complexes

Thermoresponsive Hydrogel of Diblock Methylcellulose: Formation of Ribbonlike Supramolecular Nanostructures by Self-Assembly

This article provides detailed insight into the thermoresponsive gelation mechanism of industrially produced methylcellulose (MC), highlighting the importance of diblock structure with a hydrophobic sequence of 2,3,6-tri-<i>O</i>-methyl-glucopyranosyl units for this physicochemical property. We show herein, for the first time, that well-defined diblock MC self-assembles thermoresponsively into ribbonlike nanostructures in water. A cryogenic transmission electron microscopy (cryo-TEM) technique was used to detect the ribbonlike nanostructures formed by the diblock copolymers consisting of hydrophilic glucosyl or cellobiosyl and hydrophobic 2,3,6-tri-<i>O</i>-methyl-cellulosyl blocks, methyl β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>1</b> (G-236MC, DP_<i>n</i> = 10.7, DS = 2.65), and methyl β-d-glucopyranosyl-(1→4)-β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>2</b> (GG-236MC, DP_<i>n</i> = 28.2, DS = 2.75). Rheological measurements revealed that the gel strength of a dispersion of GG-236MC (<b>2</b>, 2.0 wt %) in water at 70 °C was 3.0 times stronger than that of commercial MC SM-8000, although the molecular weight of GG-236MC (<b>2</b>) having <i>M</i>_w = 8 × 10³ g/mol was 50 times smaller than that of SM-8000 having <i>M</i>_w = 4 × 10⁵ g/mol. Cryo-TEM observation suggested that the hydrogel formation of the diblock copolymers could be attributed to the entanglement of ribbonlike nanostructures self-assembled by the diblock copolymers in water. The cryo-TEM micrograph of GG-236MC (<b>2</b>) at 5 °C showed rectangularly shaped nanostructures having a thickness from 11 to 24 nm, although G-236MC (<b>1</b>) at 20 °C showed no distinct self-assembled nanostructures. The ribbonlike nanostructures of GG-236MC (<b>2</b>) having a length ranging from 91 to 864 nm and a thickness from 8.5 to 27.1 nm were detected above 20 °C. Small-angle X-ray scattering measurements suggested that the ribbonlike nanostructures of GG-236MC (<b>2</b>) consisted of a bilayer structure with a width of ca. 40 nm. It was likely that GG-236MC (<b>2</b>) molecules were oriented perpendicularly to the long axis of the ribbonlike nanostructure. In addition, wide-angle X-ray scattering measurements revealed that GG-236MC (<b>2</b>) in its hydrogel formed the same crystalline regions as 2,3,6-tri-<i>O</i>-methylcellulose. The influence of the DP of diblock MC with a DS of around 2.7 on the gelation behavior will be discussed

Thermoresponsive Hydrogel of Diblock Methylcellulose: Formation of Ribbonlike Supramolecular Nanostructures by Self-Assembly

This article provides detailed insight into the thermoresponsive gelation mechanism of industrially produced methylcellulose (MC), highlighting the importance of diblock structure with a hydrophobic sequence of 2,3,6-tri-<i>O</i>-methyl-glucopyranosyl units for this physicochemical property. We show herein, for the first time, that well-defined diblock MC self-assembles thermoresponsively into ribbonlike nanostructures in water. A cryogenic transmission electron microscopy (cryo-TEM) technique was used to detect the ribbonlike nanostructures formed by the diblock copolymers consisting of hydrophilic glucosyl or cellobiosyl and hydrophobic 2,3,6-tri-<i>O</i>-methyl-cellulosyl blocks, methyl β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>1</b> (G-236MC, DP_<i>n</i> = 10.7, DS = 2.65), and methyl β-d-glucopyranosyl-(1→4)-β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>2</b> (GG-236MC, DP_<i>n</i> = 28.2, DS = 2.75). Rheological measurements revealed that the gel strength of a dispersion of GG-236MC (<b>2</b>, 2.0 wt %) in water at 70 °C was 3.0 times stronger than that of commercial MC SM-8000, although the molecular weight of GG-236MC (<b>2</b>) having <i>M</i>_w = 8 × 10³ g/mol was 50 times smaller than that of SM-8000 having <i>M</i>_w = 4 × 10⁵ g/mol. Cryo-TEM observation suggested that the hydrogel formation of the diblock copolymers could be attributed to the entanglement of ribbonlike nanostructures self-assembled by the diblock copolymers in water. The cryo-TEM micrograph of GG-236MC (<b>2</b>) at 5 °C showed rectangularly shaped nanostructures having a thickness from 11 to 24 nm, although G-236MC (<b>1</b>) at 20 °C showed no distinct self-assembled nanostructures. The ribbonlike nanostructures of GG-236MC (<b>2</b>) having a length ranging from 91 to 864 nm and a thickness from 8.5 to 27.1 nm were detected above 20 °C. Small-angle X-ray scattering measurements suggested that the ribbonlike nanostructures of GG-236MC (<b>2</b>) consisted of a bilayer structure with a width of ca. 40 nm. It was likely that GG-236MC (<b>2</b>) molecules were oriented perpendicularly to the long axis of the ribbonlike nanostructure. In addition, wide-angle X-ray scattering measurements revealed that GG-236MC (<b>2</b>) in its hydrogel formed the same crystalline regions as 2,3,6-tri-<i>O</i>-methylcellulose. The influence of the DP of diblock MC with a DS of around 2.7 on the gelation behavior will be discussed

Thermoresponsive Hydrogel of Diblock Methylcellulose: Formation of Ribbonlike Supramolecular Nanostructures by Self-Assembly

This article provides detailed insight into the thermoresponsive gelation mechanism of industrially produced methylcellulose (MC), highlighting the importance of diblock structure with a hydrophobic sequence of 2,3,6-tri-<i>O</i>-methyl-glucopyranosyl units for this physicochemical property. We show herein, for the first time, that well-defined diblock MC self-assembles thermoresponsively into ribbonlike nanostructures in water. A cryogenic transmission electron microscopy (cryo-TEM) technique was used to detect the ribbonlike nanostructures formed by the diblock copolymers consisting of hydrophilic glucosyl or cellobiosyl and hydrophobic 2,3,6-tri-<i>O</i>-methyl-cellulosyl blocks, methyl β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>1</b> (G-236MC, DP_<i>n</i> = 10.7, DS = 2.65), and methyl β-d-glucopyranosyl-(1→4)-β-d-glucopyranosyl-(1→4)-2,3,6-tri-<i>O</i>-methyl-celluloside <b>2</b> (GG-236MC, DP_<i>n</i> = 28.2, DS = 2.75). Rheological measurements revealed that the gel strength of a dispersion of GG-236MC (<b>2</b>, 2.0 wt %) in water at 70 °C was 3.0 times stronger than that of commercial MC SM-8000, although the molecular weight of GG-236MC (<b>2</b>) having <i>M</i>_w = 8 × 10³ g/mol was 50 times smaller than that of SM-8000 having <i>M</i>_w = 4 × 10⁵ g/mol. Cryo-TEM observation suggested that the hydrogel formation of the diblock copolymers could be attributed to the entanglement of ribbonlike nanostructures self-assembled by the diblock copolymers in water. The cryo-TEM micrograph of GG-236MC (<b>2</b>) at 5 °C showed rectangularly shaped nanostructures having a thickness from 11 to 24 nm, although G-236MC (<b>1</b>) at 20 °C showed no distinct self-assembled nanostructures. The ribbonlike nanostructures of GG-236MC (<b>2</b>) having a length ranging from 91 to 864 nm and a thickness from 8.5 to 27.1 nm were detected above 20 °C. Small-angle X-ray scattering measurements suggested that the ribbonlike nanostructures of GG-236MC (<b>2</b>) consisted of a bilayer structure with a width of ca. 40 nm. It was likely that GG-236MC (<b>2</b>) molecules were oriented perpendicularly to the long axis of the ribbonlike nanostructure. In addition, wide-angle X-ray scattering measurements revealed that GG-236MC (<b>2</b>) in its hydrogel formed the same crystalline regions as 2,3,6-tri-<i>O</i>-methylcellulose. The influence of the DP of diblock MC with a DS of around 2.7 on the gelation behavior will be discussed

Characteristics and Prognostic Impact of Pneumonitis during Systemic Anti-Cancer Therapy in Patients with Advanced Non-Small-Cell Lung Cancer - Fig 2

<p>Kaplan–Meier overall survival curves after the diagnosis of NSCLC in all patients with or without pneumonitis during systemic anti-cancer therapy (pneumonitis) (Fig 2A) and those who did not have pre-existing ILD with or without pneumonitis (Fig 2B), and survival time after the onset of pneumonitis in survivors of pneumonitis (Fig 2C).</p