Microstructural investigation of morphology and kinetics of methane hydrate in the presence of tetrabutylammonium bromide: Insights for preservation and inhibition.

Developing highly efficient methane (CH4) hydrate storage methods and understanding the hydrate dissociation kinetics can contribute to advancing CH4 gas storage and transport. The effects of tetrabutylammonium bromide (TBAB) (a thermodynamic promoter) addition on the kinetics of CH4 hydrate were evaluated on the microscopic scale using synchrotron x-ray computed tomography (CT) and powder x-ray diffraction. Microscopic observations showed that a 5 wt. % TBAB solution facilitated the nucleation of CH4 hydrate owing to the initial growth of TBAB semi-clathrate hydrate particles. The CH4 hydrate crystals in the CH4 + TBAB hydrate sample were sponge-like with many internal pores and exhibited slightly enhanced self-preservation compared to the pure CH4 hydrate, both in the bulk and after pulverization to a fine powder. This study demonstrates the feasibility of controlling the rate of CH4 hydrate formation and preservation by using aqueous TBAB solutions in CH4 hydrate formation.

Multi-phase retrieval of methane hydrate in natural sediments by cryogenic x-ray computed tomography.

In this study, we observed natural methane (CH4) hydrate sediments, which are a type of unconventional natural gas resources, using x-ray computed tomography (CT). Because CH4 hydrates are formed by hydrogen bonding of water molecules with CH4, material decomposition becomes challenging when CH4 hydrates coexist with liquid or solid water in natural sediments. Tri-contrast (absorption, refraction, and scattering) imaging was performed via diffraction enhanced x-ray CT optics using monochromatic synchrotron x rays. The quantitative characterization of the contrast changes successfully enabled the decomposition of CH4 hydrates coexisting with frozen seawater (ice) in natural sediments obtained from the Okhotsk Sea. This study reveals complementary structural information about the microtexture and spatial relation among CH4 hydrates, ice, and pores by utilizing the distinct physical properties of x rays when passing through the materials. These results highlight the exceptional capabilities of high-resolution multicontrast x-ray tomography in materials science and geoscience applications.

BN-Embedded Aromatic Hydrocarbon Synthesis via Nucleophilic Diboration Reactions.

Activation of bis(pinacolato)diboron with aromatic lithium amide promotes diboration of the proximal C-C triple bond, leading to BN-embedded aromatic compounds. In situ treatment of the initially generated spirocyclic borate intermediate with aqueous acid or organometallic reagents enables ligand installation on the endocyclic boron atom.

A gold-complex initiated functionalization of biologically active polyphenols applied to a 18F-labeled chemical probe.

(-)-Epigallocatechin gallate (EGCG), a key component of green tea, exerts therapeutic anticancer and antiallergic properties through its binding to the 67 kDa laminin receptor. The functionalization of EGCG is a promising strategy for creating new drug candidates and chemical probes. In our study, we developed a method for effectively modifying the A ring of EGCG through an electrophilic aromatic substitution with amidomethyl 2-alkynylbenzoates initiated with a gold complex. The 2-alkynylbenzoates treated with (Ph3P)AuOTf under neutral conditions yielded N-acylimines. A further electrophilic aromatic substitution resulted in a mixture of EGCG substituted with acylaminomethyl groups at the 6 and 8 positions with a significant amount noted at the 6 position. We then explored the synthesis of 18F-labeled EGCG with a neopentyl labeling group, an effective labeling group for radiohalogens of not only fluorine-18 but also of astatine-211. To achieve this, we prepared precursors that possessed acid-sensitive protecting groups and base-unstable leaving groups using our established method. Substitution of EGCG with a neopentyl labeling group at either the C6 or C8 position did not affect its anticancer efficacy in U266 cells. Finally, we investigated the preparation of 18F-labeled EGCG. The 18F-fluorination of a mixture of 6- and 8-substituted precursors yielded the corresponding 18F-labeled compounds in 4.5% and 3.0% radiochemical yields (RCYs), respectively. Under acidic conditions, the 18F-labeled 8-substituted compound produced 18F-labeled EGCG in 37% RCY, which heralds the potential of our functionalization approach.

Perfluoroalkoxylation reaction via dual concurrent catalysis.

A catalytic amount of CsI enables dual concurrent activation of poorly reactive perfluoroalkoxide and alkyl halides, especially alkyl chlorides, leading to the formation of diverse perfluoroalkoxylated organic compounds. Installation of perfluoroalkoxy groups by this methodology is cost-effective, circumventing the need for over-stoichiometric cesium or silver salts. This methodology also provides high functional group compatibility and tolerance of sterically hindered substrates.

Alignment and use of microbeam with full-field x-ray microscopes.

Demonstration tests of the alignment of Fresnel zone plate focusing optics using a full-field x-ray microscope and microbeam x-ray diffraction measurements combined with the full-field x-ray microscope were performed. It was confirmed that the full-field x-ray microscope enables direct two-dimensional observation of a microbeam with sub-micrometer spatial resolution. This allowed visualization of the misalignment of the focusing optics, resulting in accurate alignment of the optics within a short time. In addition, the microscope could be used to observe the sample as well as the microbeam, which enabled clarification of the position and two-dimensional shape of the microbeam on the sample. This realized a measurement procedure that a 100-µm-size sample was imaged with sub-micrometer spatial resolution, and then, microbeam-use measurements were performed for only the region of interest determined by the microscope, which has been difficult with conventional microbeam applications. The combination of observations by a full-field x-ray microscope and measurements using a microbeam is expected to open a new style of measurement.

Correction: Visualizing morphological structures of rice grains in precooked products using synchrotron radiation X-ray phase-contrast computed tomography.

Correction for 'Visualizing morphological structures of rice grains in precooked products using synchrotron radiation X-ray phase-contrast computed tomography' by Hiromi Miki et al., Food Funct., 2023, 14, 87-93, https://doi.org/10.1039/D2FO02714C.

Visualizing morphological structures of rice grains in precooked products using synchrotron radiation X-ray phase-contrast computed tomography.

Rice is a staple food eaten by more than half of the world's population. In recent years, various types of precooked rice products have become commercially available as more and more people, including the elderly, do not have time to prepare meals or are coping with eating disorders. To address this situation, analysis of the internal structure of rice is important to understand the phenomena that occur during cooking rice grains and to develop rice products for specific purpose with desired physical properties. Although the internal structure of food has been observed by many methods, it is difficult to image the internal structure of rice porridge or rice grains with sufficient spatial resolution in a nondestructive manner. In this study, we applied X-ray interferometer-based phase-contrast computed tomography for the nondestructive observation of precooked rice products. For comparison, X-ray absorption-contrast images were also observed. The samples consisted of packaged aseptic cooked rice, precooked rice porridge for the general public, and a texture-modified rice product for people with dysphagia. In the phase-contrast two-dimensional (2D) tomographic images, the central lines of the cooked rice grains were successfully observed in all samples. The changes in the peripheral regions and internal hollows of the cooked rice grains were also clearly observed. Meanwhile, rice grains were hardly discernible in the X-ray absorption images. These results indicate that X-ray interferometer-based phase-contrast imaging is a promising tool for analyzing the morphology and internal structure of cooked rice grains in various prepacked products such as rice porridge and texture-modified rice.

Practical and Facile Access to Bicyclo[3.1.1]heptanes: Potent Bioisosteres of meta-Substituted Benzenes.

There is increasing interest in replacement of the planar aromatic rings of drug candidates with three-dimensional caged scaffolds in order to improve the physical properties, but bioisosteres of meta-substituted benzenes have remained elusive. We focused on the bicyclo[3.1.1]heptane (BCH) scaffold as a novel bioisostere of meta-substituted benzenes, anticipating that [3.1.1]propellane (2) would be a versatile precursor of diversely functionalized BCHs. Here, we describe a practical preparative method for [3.1.1]propellane from newly developed 1,5-diiodobicyclo[3.1.1]heptane (1), as well as difunctionalization reactions of 2 leading to functionalized BCHs. We also report postfunctionalization reactions of these products.

Cardio-Ankle Vascular Index Reflects the Efficacy of Waon Therapy in Heart Failure Patients.

Waon therapy is a form of thermal treatment in a dry sauna developed by Tei. Although Waon therapy is reportedly effective for chronic heart failure (CHF) patients, not all patients respond to the therapy. The reason for this ineffectiveness has not been fully clarified. The cardio-ankle vascular index (CAVI) is an index of arterial stiffness of the arterial tree from the origin of the aorta to the ankle, and it is thought to reflect some of the afterload of the left ventricle. We investigated the effects of Waon therapy on CAVI and plasma brain natriuretic peptide (BNP) level to clarify the usefulness of CAVI during Waon therapy.CHF patients (n = 21) treated with Waon therapy (2 weeks of 10 sessions) were divided into two groups: responders with an improved BNP level (n = 11) and nonresponders with no improvement in BNP (n = 10). CAVI was measured using Vasela 1500.A significant decrease in CAVI (median and interquartile range) was observed in the responder group (from 10.3 [9.6, 11.6] to 9.6 [8.6, 10.3], P = 0.021), whereas no change was observed in the nonresponder group (from 9.6 [8.6, 10.5] to 9.5 [9.1, 11.2], P = 0.919). The incidence of rehospitalization or cardiac death due to heart failure was significantly higher in patients in whom Waon therapy was ineffective at 12 months of follow-up (log-rank P = 0.001).The effectiveness of Waon therapy in CHF patients may be reflected by the improvement in CAVI.

X-ray zooming optics for analyzer-based multi-contrast computed tomography.

An X-ray analyzer-based optics with a zoom function is proposed for observing various samples with apparent-absorption contrast, phase contrast and scattering contrast. The proposed X-ray optics consists of a collimator crystal and an analyzer crystal arranged in a nondispersive (+, -) geometry with a sample placed between them. For the implementation of the zoom function, an asymmetrically cut crystal in the rotated-inclined geometry was used for the analyzer. Proof-of-principle experiments were performed at the vertical wiggler beamline BL-14B of the Photon Factory. First, the magnification was set to 1×, and then it was zoomed into the optimal magnification (10×). At these magnifications, tri-modal contrast cross-sectional images of a sample were obtained by computed tomography. It was confirmed that the image quality at 10× was superior to that at 1×. This achievement opens up new possibilities for observing an entire sample or regions of interest within a sample at optimal magnification, and is expected to be useful for materials science, condensed matter physics, archeology and biomedical science.

Existence of twisting in dislocation-free protein single crystals.

The growth of high-quality protein crystals is a prerequisite for the structure analysis of proteins by X-ray diffraction. However, dislocation-free perfect crystals such as silicon and diamond have been so far limited to only two kinds of protein crystals, such as glucose isomerase and ferritin crystals. It is expected that many other high-quality or dislocation-free protein crystals still exhibit some imperfection. The clarification of the cause of imperfection is essential for the improvement of crystallinity. Here, we explore twisting as a cause of the imperfection in high-quality protein crystals of hen egg-white lysozyme crystals with polymorphisms (different crystal forms) by digital X-ray topography with synchrotron radiation. The magnitude of the observed twisting is 10−6 to 10−5°/µm which is more than two orders smaller than 10−3 to 104°/µm in other twisted crystals owing to technique limitations with optical and electron microscopy. Twisting is clearly observed in small crystals or in the initial stage of crystal growth. It is uniformly relaxed with crystal growth and becomes smaller in larger crystals. Twisting is one of main residual defects in high-quality crystals and determines the crystal perfection. Furthermore, it is presumed that the handedness of twisting can be ascribed to the anisotropic interaction of chiral protein molecules associated with asymmetric units in the crystal forms. This mechanism of twisting may correspond to the geometric frustration proposed as a primary mechanism of twisting in molecular crystals. Our finding provides insights for the understanding of growth mechanism and the growth control of high-quality crystals.

Observation of processed rice using synchrotron radiation X-ray phase-contrast imaging.

As a result of increasing elderly population, the number of people with dysphagia and difficulty eating is expected to increase as well. It is not always feasible to cook meals to the proper texture for elderly individuals and people with dysphagia living at home. Accordingly, there is high market demand for ready-made texture-modified foods for home consumption. Many methods have been widely used to analyze the structural properties of foods. However, it is difficult to observe the structure of foods in a nondestructive manner with the conventional analysis methods. To observe food as it is without destroying it, we adopted X-ray phase-contrast computed tomography (CT) using a crystal interferometer in this study. The samples were commercially available precooked rice porridge and two texture-modified rice products satisfying the Universal Design Food criteria. A texture profile analysis was also performed to compare the phase-contrast images and the food properties. In the phase-contrast images, both the ingredients and cohesion in the samples were successfully observed. It was also found that the texture-modified rice products were easier to eat and swallow than the rice porridge. These results show that X-ray phase-contrast CT using a crystal interferometer is a promising method for nondestructive observation of food structures and is expected to open up new horizons in food science.

X-ray zooming microscopy with two Fresnel zone plates.

We propose a variable-magnification full-field x-ray microscope using two Fresnel zone plates (FZPs). By moving the positions of the two FZPs, the magnification can be continuously changed even if the sample and camera positions are fixed. It was demonstrated that the magnification can be changed in the range of 25-150× using a hard x-ray beam at 14.4 keV. Using the first FZP as a convex lens and the second FZP as a concave lens, high magnification can be achieved at a short camera length. Even under the condition of a camera length of about 7 m, a magnification higher than 300× was achieved, and a line and space pattern with a pitch of 40 nm was observed at 10 keV. By inserting a knife edge at an appropriate position in the optical system, a phase-contrast image can be easily obtained, which is useful for soft-tissue observation of biological samples.

Superheating of Structure I Gas Hydrates within the Structure II Cyclopentane Hydrate Shell.

The superheated state of methane (CH4) hydrate that exists under the surface ice layer can persist for considerable lengths of time, which showed promise as a method for storing and transporting natural gas. This study extends this further by coating sI CH4 hydrate with one of several sII hydrates, thus eliminating the need for a defect-free continuous interface between the sI and sII hydrates. Gas hydrate crystals were kept intact above their dissociation temperature by immersing them in liquid cyclopentane (CP), as observed with powder X-ray diffraction and X-ray CT methods. It was observed that placing the CH4 hydrate in CP converted the outer layer of CH4 hydrate to a thin layer of CP hydrate at around 270 K under atmospheric pressure, which is ∼80 K higher than the usual dissociation temperature. It was also observed that sI CO2 hydrate and C2H6 hydrate could be preserved by CP hydrate.

Nucleophilic Activation of Hydrosilanes via a Strain-Imposing Strategy Leading to Functional Sila-aromatics.

Carefully designed cyclic hydrosilanes enable trans-selective hydrosilylation of unactivated alkynes without transition metal catalysts via silicate formation. Employment of sterically demanding bidentate ligands of silicon increases steric congestion upon silicate formation, and this strain-imposing strategy facilitates hydride transfer. This hydrosilylation provides efficient access to diverse benzosiloles, silaphenalenes, and related silacycles.

Role of the N- and C-terminal regions of FliF, the MS ring component in Vibrio flagellar basal body.

The MS ring is a part of the flagellar basal body and formed by 34 subunits of FliF, which consists of a large periplasmic region and two transmembrane segments connected to the N- and C-terminal regions facing the cytoplasm. A cytoplasmic protein, FlhF, which determines the position and number of the basal body, supports MS ring formation in the membrane in Vibrio species. In this study, we constructed FliF deletion mutants that lack 30 or 50 residues from the N-terminus (ΔN30 and ΔN50), and 83 (ΔC83) or 110 residues (ΔC110) at the C-terminus. The N-terminal deletions were functional and conferred motility of Vibrio cells, whereas the C-terminal deletions were nonfunctional. The mutants were expressed in Escherichia coli to determine whether an MS ring could still be assembled. When co-expressing ΔN30FliF or ΔN50FliF with FlhF, fewer MS rings were observed than with the expression of wild-type FliF, in the MS ring fraction, suggesting that the N-terminus interacts with FlhF. MS ring formation is probably inefficient without FlhF. The deletion of the C-terminal cytoplasmic region did not affect the ability of FliF to form an MS ring because a similar number of MS rings were observed for ΔC83FliF as with wild-type FliF, although further deletion of the second transmembrane segment (ΔC110FliF) abolished it. These results suggest that the terminal regions of FliF have distinct roles; the N-terminal region for efficient MS ring formation and the C-terminal region for MS ring function. The second transmembrane segment is indispensable for MS ring assembly.ImportanceThe bacterial flagellum is a supramolecular architecture involved in cell motility. At the base of the flagella, a rotary motor that begins to construct an MS ring in the cytoplasmic membrane comprises 34 transmembrane proteins (FliF). Here, we investigated the roles of the N and C terminal regions of FliF, which are MS rings. Unexpectedly, the cytoplasmic regions of FliF are not indispensable for the formation of the MS ring, but the N-terminus appears to assist in ring formation through recruitment of FlhF, which is essential for flagellar formation. The C-terminus is essential for motor formation or function.

Effects of 18F-fluorinated neopentyl glycol side-chain on the biological characteristics of stilbene amyloid-ß PET ligands.

INTRODUCTION: The 2,2-dihydroxymethyl-1-[18F]fluoropropane group, also called 18F-labelled neopentyl glycol side-chain, is a novel 18F-labelling group for positron emission tomography (PET) imaging agents. The aim of using this group is to develop simple purification with solid-phase extraction without high-performance liquid chromatography. However, the effects of the neopentyl 18F-labelling group on the characteristics of brain imaging agents are unknown. Here, we added this side-chain to compounds with an aminostilbene structure to evaluate their effects on the biological properties of aminostilbene as an amyloid-ß (Aß) radioligand. METHODS: Biodistributions of four novel 18F-labelled stilbene compounds with different lengths of polyethylene glycol (PEG) linkers, called [18F]Cpd-0, -1, -2, and -4, (PEG = 0, 1, 2, and 4), and [18F]AV-1 in normal mice were evaluated. Metabolite analysis of [18F]Cpd-0 and -1 was performed with mouse plasma and brain. A competitive binding assay of [18F]AV-1 binding to Aß1-42 fibrils was performed to determine the binding properties of the compounds. RESULTS: [18F]Cpd-0, -1, and -2 demonstrated moderate initial brain uptake in mice (3.1-4.2% injected dose/g at 2 min post-injection) followed by fast clearance, and in vivo defluorination of these compounds was negligible. [18F]Cpd-4 exhibited low brain uptake and high bone uptake. Compared with [18F]Cpd-1, the percentage of [18F]Cpd-0 in mouse brain was high at 10 min post-injection. A competitive binding assay revealed partial interference effects by the neopentyl glycol side-chain on binding of stilbene compounds to Aß1-42 fibrils. CONCLUSIONS: Aminostilbene compounds with two or fewer PEG linkers containing an 18F-labelled neopentyl glycol side-chain demonstrated preferable pharmacokinetic properties as a brain imaging radioligand in normal mice. These side-chains can be used as an alternative labelling group for imaging agents targeting the brain.

Nucleophilic Diboration Strategy Targeting Diversified 1-Boraphenarene Architectures.

1-Boraphenalenes, a class of boron-doped polyaromatic hydrocarbons, were synthesized by nucleophilic diboration reaction of alkynes. Activation of diboron reagents with a highly basic sp2 -carbanion results in very fast successive C-B bond formations to construct the boracycle. This methodology is characterized by high chemoselectivity, affording a wide variety of 1-boraphenarenes with diverse polar substituents. The endocyclic boron can be arylated conveniently in one pot, and the peripheral boron is available for various chemical transformations. Highly diastereoselective diboration gives pseudo-enantio-enriched boraphenarene, which emits circularly polarized fluorescence (CPL).

Assembly mechanism of a supramolecular MS-ring complex to initiate bacterial flagellar biogenesis in Vibrio species.

The bacterial flagellum is an organelle responsible for motility and has a rotary motor comprising the rotor and the stator. Flagellar biogenesis is initiated by the assembly of the MS-ring, a supramolecular complex embedded in the cytoplasmic membrane. The MS-ring consists of a few dozen copies of the transmembrane FliF protein, and is an essential core structure which is a part of the rotor. The number and location of the flagella are controlled by the FlhF and FlhG proteins in some species. However, there is no clarity on the factors initiating MS-ring assembly, and contribution of FlhF/FlhG to this process. Here, we show that FlhF and a C-ring component FliG facilitate Vibrio MS-ring formation. When Vibrio FliF alone was expressed in Escherichia coli cells, MS-ring formation rarely occurred, indicating the requirement of other factors for MS-ring assembly. Consequently, we investigated if FlhF aided FliF in MS-ring assembly. We found that FlhF allowed GFP-fused FliF to localize at the cell pole in a Vibrio cell, suggesting that it increases local concentration of FliF at the pole. When FliF was co-expressed with FlhF in E. coli cells, the MS-ring was effectively formed, indicating that FlhF somehow contributes to MS-ring formation. The isolated MS-ring structure was similar to the MS-ring formed by Salmonella FliF. Interestingly, FliG facilitates MS-ring formation, suggesting that FliF and FliG assist in each other's assembly into the MS-ring and C-ring. This study aids in understanding the mechanism behind MS-ring assembly using appropriate spatial/temporal regulations.Importance Flagellar formation is initiated by the assembly of the FliF protein into the MS-ring complex, embedded in the cytoplasmic membrane. The appropriate spatial/temporal control of MS-ring formation is important for the morphogenesis of the bacterial flagellum. Here, we focus on the assembly mechanism of Vibrio FliF into the MS-ring. FlhF, a positive regulator of the number and location of flagella, recruits the FliF molecules at the cell pole and facilitates MS-ring formation. FliG also facilitates MS-ring formation. Our study showed that these factors control flagellar biogenesis in Vibrio, by initiating the MS-ring assembly. Furthermore, it also implies that flagellar biogenesis is a sophisticated system linked with the expression of certain genes, protein localization and a supramolecular complex assembly.