Trace element mapping of a single cell using a hard x‐ray nanobeam focused by a Kirkpatrick‐Baez mirror system

To visualize the distributions of trace elements in biological samples such as tissues and cells at high spatial resolution, we developed a scanning x‐ray fluorescence microscope (SXFM) at SPring‐8, using a Kirkpatrick‐Baez mirror optics that enables achromatic and highly efficient focusing. To evaluate performance regarding its application to biological samples, the SXFM was used at x‐ray energy of 15 keV to observe NIH/3T3 cells in which adenosine triphosphate (ATP) synthase β (specifically localized at the mitochondria) were labeled with gold colloidal particles. Various elemental distributions were visualized at the single‐cell level, including those for P, S, Cl, Ca, Fe, Cu, Zn and Au, and we obtained high‐resolution elemental distribution maps by magnifying the labeled single mitochondrion. Maximum spatial resolution achieved in the experiments was sub‐100 nm. Copyright © 2008 John Wiley & Sons, Ltd.     

Hypervalent iodine(III)/Et4NBr combination in water for green and racemization-free aqueous oxidation of alcohols

We have found that the use of the PhI(OAc)₂/Et₄N⁺Br⁻ combination in water can significantly enhance its oxidation ability and oxidize a wide range of alcohols 1 to carbonyl compounds 2 in good to excellent yields. This clean aqueous oxidation method shows no detectable racemization processes, and even an enolizable ketone 2m could be obtained in an optically pure form from the corresponding chiral alcohol 1m. Utilization of the recyclable reagent 3 as a more practical alternative to PhI(OAc)₂ is also successful in these reactions.     

Cation-pi interaction in the polyolefin cyclization cascade uncovered by incorporating unnatural amino acids into the catalytic sites of squalene cyclase

It has been assumed that the pi-electrons of aromatic residues in the catalytic sites of triterpene cyclases stabilize the cationic intermediates formed during the polycyclization cascade of squalene or oxidosqualene, but no definitive experimental evidence has been given. To validate this cation-pi interaction, natural and unnatural aromatic amino acids were site-specifically incorporated into squalene-hopene cyclase (SHC) from Alicyclobacillus acidocaldarius and the kinetic data of the mutants were compared with that of the wild-type SHC. The catalytic sites of Phe365 and Phe605 were substituted with O-methyltyrosine, tyrosine, and tryptophan, which have higher cation-pi binding energies than phenylalanine. These replacements actually increased the SHC activity at low temperature, but decreased the activity at high temperature, as compared with the wild-type SHC. This decreased activity is due to the disorganization of the protein architecture caused by the introduction of the amino acids more bulky than phenylalanine. Then, mono-, di-, and trifluorophenylalanines were incorporated at positions 365 and 605; these amino acids reduce cation-pi binding energies but have van der Waals radii similar to that of phenylalanine. The activities of the SHC variants with fluorophenylalanines were found to be inversely proportional to the number of the fluorine atoms on the aromatic ring and clearly correlated with the cation-pi binding energies of the ring moiety. No serious structural alteration was observed for these variants even at high temperature. These results unambiguously show that the pi-electron density of residues 365 and 605 has a crucial role for the efficient polycyclization reaction by SHC. This is the first report to demonstrate experimentally the involvement of cation-pi interaction in triterpene biosynthesis.     

Intersite structural rearrangement of the blue copper site induced by substrate binding: spectroscopic studies of a copper-containing nitrite reductase from Alcaligenes xylosoxidans NCIMB 11015

A copper-containing nitrite reductase from Alcaligenes xylosoxidans NCIMB 11015 has its own unique blue or type 1 copper protein resonance Raman spectrum in the usual Cu-S(Cys) stretching region, nu(Cu-S(Cys)), with a pair of strong peaks at 412 and 420 cm(-1) and a weak peak at 364 cm(-1). The predominantly nu(Cu-S(Cys)) Raman bands at 412, 420, and 364 cm(-1) of the type 1 copper site all shifted to higher frequencies upon binding of nitrite to the type 2 copper site, and the resonance Raman difference spectra progressively intensified with the increments of nitrite ion concentration. Positive support for substrate binding to the type 2 copper is provided by the nu(Cu-S(Cys)) bands in the resonance Raman spectrum of a type 2 copper-depleted enzyme, which is insensitive to the presence of NO2-. The shift to higher frequency of the Raman bands of the type 1 copper center with the addition of nitrite ions suggests a stronger Cu-S(Cys) interaction in the substrate-bound A. xylosoxidans nitrite reductase.     

Modified Vonk's Method to Determine Crystallinity and Crystal Distortion in Polymers

Using a modified method developed from Vonk's method, detailed values of crystallinity and crystal disorder were obtained by wide angle X-ray diffraction (WAXD). In Vonk's method, the crystallinity (w) is determined by extrapolation of the WAXD experimental curve back to zero scattering angle, while the distortion factor (k) is determined by the inclination of the experimental curve at zero scattering angle. In our new method, both w and k are determined at the same time by using the least squares method. In order to show the efficiency of our method, the new fitting procedure was applied to the experimental values of polyethylene and polyethylene terephthalate as measured by Vonk, and the values of w and k determined by our new method were compared with those found by Vonk's method. The new fitting method has the advantage that it uses the whole experimental curve. Moreover, our modified Vonk's method enables us to discuss crystal distortions as arising from first-kind (thermal) disorder and second-kind (paracrystalline) disorder.     

Comparison of accuracy of intravoxel incoherent motion and apparent diffusion coefficient techniques for predicting malignancy of head and neck tumors using half-Fourier single-shot turbo spin-echo diffusion-weighted imaging

Purpose

To evaluate the use of the intravoxel incoherent motion (IVIM) technique in half-Fourier single-shot turbo spin-echo (HASTE) diffusion-weighted imaging (DWI), and to compare its accuracy to that of apparent diffusion coefficient (ADC) to predict malignancy in head and neck tumors.

Patients and methods

HASTE DW images of 33 patients with head and neck tumors (10 benign and 23 malignant) were evaluated. Using the IVIM technique, parameters (D, true diffusion coefficient; f, perfusion fraction; D*, pseudodiffusion coefficient) were calculated for each tumor. ADC values were measured over a range of b values from 0 to 1000 s/mm². IVIM parameters and ADC values in benign and malignant tumors were compared using Student's t test, receiver operating characteristics (ROC) analysis, and multivariate logistic regression modeling.

Results

Mean ADC and D values of malignant tumors were significantly lower than those of benign tumors (P < 0.05). Mean D* values of malignant tumors were significantly higher than those of benign tumors (P < 0.05). There was no significant difference in mean f values between malignant and benign tumors (P > 0.05). The technique of combining D and D* was the best for predicting malignancy; accuracy for this model was higher than that for ADC.

Conclusions

The IVIM technique may be applied in HASTE DWI as a diagnostic tool to predict malignancy in head and neck masses. The use of D and D* in combination increases the diagnostic accuracy in comparison with ADC.     

Hydrothermal and solid-state transformation of ruthenium-supported Keggin-type heteropolytungstates [XW11O39{Ru(II)(benzene)(H2O)}]n- (X = P (n = 5), Si (n = 6), Ge (n = 6)) to ruthenium-substituted Keggin-type heteropolytungstates

A new pathway for the preparation of mono-ruthenium (Ru)(iii)-substituted Keggin-type heteropolytungstates with an aqua ligand, [PW(11)O(39)Ru(iii)(H(2)O)](4-) (1a), [SiW(11)O(39)Ru(iii)(H(2)O)](5-) (1b) and [GeW(11)O(39)Ru(iii)(H(2)O)](5-) (1c), using [Ru(ii)(benzene)Cl(2)](2) as a Ru source was described. Compounds 1a-1c were prepared by reacting [XW(11)O(39)](n-) (X = P, Si and Ge) with [Ru(ii)(benzene)Cl(2)](2) under hydrothermal condition and were isolated as caesium salts. Ru(benzene)-supported heteropolytungstates, [PW(11)O(39){Ru(ii)(benzene)(H(2)O)}](5-) (2a), [SiW(11)O(39){Ru(ii)(benzene)(H(2)O)}](6-) (2b) and [GeW(11)O(39){Ru(ii)(benzene)(H(2)O)}](6-) (2c), were first produced in the reaction media, and then transformed to 1a, 1b and 1c, respectively, under hydrothermal conditions. Calcination of Ru(benzene)-supported heteropolytungstates, 2a, 2b and 2c, in the solid state produced mixtures of 1a, 1b and 1c with CO (carbon monoxide)-coordinated complexes, [PW(11)O(39)Ru(ii)(CO)](5-) (4a), [SiW(11)O(39)Ru(ii)(CO)](6-) (4b) and [GeW(11)O(39)Ru(ii)(CO)](6-) (4c), respectively. From comparison of their catalytic activities in water oxidation reaction, it was indicated that ruthenium should be incorporated in the heteropolytungstate in order to promote catalytic activity.     

Reduction of thermal fluctuations in a cryogenic laser interferometric gravitational wave detector

The thermal fluctuation of mirror surfaces is the fundamental limitation for interferometric gravitational wave (GW) detectors. Here, we experimentally demonstrate for the first time a reduction in a mirror's thermal fluctuation in a GW detector with sapphire mirrors from the Cryogenic Laser Interferometer Observatory at 17 and 18 K. The detector sensitivity, which was limited by the mirror's thermal fluctuation at room temperature, was improved in the frequency range of 90 to 240 Hz by cooling the mirrors. The improved sensitivity reached a maximum of 2.2×10(-19) m/√Hz at 165 Hz.     

Molecular recognitive adsorption of aqueous tetramethylammonium on the organic derivative of Hiroshima University Silicate-1 with a silane coupling reagent

Hiroshima University Silicate-1 (HUS-1), composed of silicate sheets with a halved sodalite cage and the interlayer tetramethylammonium (TMA) cation in the cage, was modified with dimethyldichlorosilane to form the organic derivative in which a dimethyl group was grafted onto the interlayer surface and a part of the TMA was removed, and the silylated HUS-1 effectively and selectively adsorbed TMA from water even in the presence of aqueous phenol.     

Preparation of Co–Al and Ni–Al layered double hydroxide thin films by a sol–gel process with hot water treatment

Using hot water treatment of sol–gel derived precursor gel films, Co–Al and Ni–Al layered double hydroxide (LDH) thin films were prepared. The precursor gel films of Al₂O₃–CoO or Al₂O₃–NiO were prepared from cobalt or nickel nitrates and aluminum tri-sec-butoxide using the sol–gel method. Then, the precursor gel films were immersed in a NaOH aqueous solution of 100 °C. Nanocrystallites of Co–Al and Ni–Al LDH were precipitated with the hot water treatment with NaOH solution. The largest amounts of nanocrystals were obtained with a solution of pH = 10 for Co–Al LDH, and with that of pH = 9 for Ni–Al LDH. X-ray diffraction measurements confirmed that this process formed CO₃ ²⁻ intercalated LDHs. Both Co–Al and Ni–Al LDH thin films were confirmed to work as electrodes for electrochemical devices by cyclic voltammogram measurements.