Identification of Shiga toxins in Shiga toxin-producing Escherichia coli using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry

We developed a rapid and reliable identification method for Shiga toxins in Shiga toxin-producing Escherichia coli (STEC) using immunoprecipitation and high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). Polyclonal antisera specific for Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) were raised in rabbits so as to be used for the immunoprecipitation. The immunoprecipitaion was carried out by mixing sample solutions with 50 microl each of the antisera to Stx1 and Stx2 followed by allowing the mixed solutions to stand for 30 min. The quantity required to obtain the immunoprecipitate was more than 0.5 microg of Shiga toxins. HPLC-ESI-MS analysis of the resulting immunoprecipitates provided accurate molecular weight information on Shiga toxins, leading to direct evidence for the presence of these toxins. It requires at most two days to perform our procedure from toxin extraction to measurement of HPLC-ESI-MS whereas the previous method using isolation procedures required about two weeks to complete. The usefulness of the present method has been demonstrated by identifying Stx1, Stx2 and a variant of Stx2 (Stx2e) in the immunoprecipitates prepared from STEC strains.     

Total synthesis of merrilactone A

Merrilactone A (1) has been shown to possess neurotrophic activity and thus is expected to hold therapeutic potential in the treatment of neurodegeneration diseases. In this paper, we report the total synthesis of (+/-)-1, employing, as key steps, a novel desymmetrization protocol of meso-diketone to construct the core cis-bicyclo[3.3.0]octyl system of 1 (3 --> 2) and a radical cyclization to install the highly congested C9-quaternary carbon (16 --> 17).     

Multicolor photochromism of two- and three-component diarylethene crystals

Novel photochromic single crystals composed of three different kinds of diarylethenes, 1,2-bis(3,5-dimethyl-2-thienyl)perfluorocyclopentene (1a), 1,2-bis(2,5-dimethyl-3-thienyl)perfluorocyclopentene (2a), and 1,2-bis(2-methyl-5-p-methoxyphenyl-3-thienyl)perfluorocyclopentene (3a), have been prepared. The three-component crystals turned yellow, orange, red, purple, blue, green, or black upon irradiation with light of appropriate wavelengths. The colors of the crystals were thermally stable in the dark and completely bleached by irradiation with visible light. Such multicolored photochromic crystals have potential for the application to optoelectronic devices, such as multifrequency three-dimensional optical memory media or full-color displays.     

Estimation of electrophoretic mobilities of red blood cells in 1-G and microgravity using a miniature capillary electrophoresis unit

Electrophoretic mobilities of red blood cells (RBCs) were measured in microgravity using a home-made capillary electrophoresis unit, which consisted of two small reservoirs of 0.6 mL and a fused-silica capillary tubing with 2 cm in length and 50 num in inner diameter. Migration of RBCs was observed by a microscope at 1000 times magnification and recorded on a videotape. The experiments were performed during stays in microgravity (about 0.01 G), which lasted 20 s and were attained by parabolic flights of an aircraft. On average, the electrophoretic mobilities of RBCs determined in microgravity were about 30% higher than those measured at 1-G condition irrespectively whether the cells were suspended in saline or serum during measurements. This difference might be explained as being mainly due to the cell floating in microgravity. Morphological changes of RBCs may contribute partly to the difference, while the variation in viscosity of the medium under microgravity could play only a minor role.     

Novel assembly of cyclic ethers by coupling alpha-chlorosulfides and alcohols

A novel protocol for assembling polycyclic ethers was developed and successfully applied to the synthesis of the EFGH ring system of ciguatoxin CTX3C. A key transformation involves construction of an O,S-acetal through coupling of alpha-chlorosulfide and a secondary alcohol under mild conditions. The method is highly applicable to use with sensitive substrates and will enable the synthesis of various natural and artificial polycyclic ethers. [reaction: see text]     

New microporous coordination polymer affording guest-coordination sites at channel walls

Utilization of a metalloligand, ([Cu(2,4-pydca)2(H2O)].2Et3NH) (1) (2,4-pydca = pyridine-2,4-dicarboxylate), as a building unit provides a novel porous coordination polymer, ([ZnCu(2,4-pydca)2(H2O)3(DMF)].DMF)n (2), in which the Zn(II) ion at the node of the network acts as a linker and the Cu(II) ion in the channel wall is available for guest-coordination.     

Construction of the benzylic quaternary carbon center of zoanthenol by intramolecular Mizoroki-Heck reaction of enone

[reaction: see text]. Stereocontrolled synthesis of the ABC ring framework of zoanthenol has been achieved. Our studies show that a beta,beta-disubstituted enone can act as a good acceptor of arylpalladium intermediates in the formation of a congested benzylic quaternary carbon center through an intramoleculer Mizoroki-Heck reaction. The cis B/C ring system was stereoselectively converted to the trans-fused framework through a SmI2-promoted deoxygenation of the alpha-hydroxy ketone.     

Dithienylethenes with a novel photochromic performance

Dithienylethenes with low decoloration quantum yields and thermal reversibility at high temperature above 100 degrees C were prepared. Introduction of bulky alkoxy substituents at 2- and 2'-positions of the thiophene rings strongly suppressed the cycloreversion quantum yields. The quantum yields were lower than 10(-3), and the photogenerated color remained stable enough under room light. On the other hand, the bulky alkoxy substituent decreased the thermal stability of the colored closed-ring isomers at high temperature. The color of the dithienylethene with cyclohexyloxy substituents faded out in less than 1 min at 160 degrees C.     

Visualizing Microglia with a Fluorescence Turn‐On Ugt1a7c Substrate

Microglia, the brain‐resident macrophage, are involved in brain development and contribute to the progression of neural disorders. Despite the importance of microglia, imaging of live microglia at a cellular resolution has been limited to transgenic mice. Efforts have therefore been dedicated to developing new methods for microglia detection and imaging. Using a thorough structure–activity relationships study, we developed CDr20, a high‐performance fluorogenic chemical probe that enables the visualization of microglia both in vitro and in vivo. Using a genome‐scale CRISPR‐Cas9 knockout screen, the UDP‐glucuronosyltransferase Ugt1a7c was identified as the target of CDr20. The glucuronidation of CDr20 by Ugt1a7c in microglia produces fluorescence.     

Recent progress on catalytic [1,3]-oxygen rearrangement reactions from nitrogen to carbon atoms

[1,3]-Rearrangement reactions of an oxygen atom from a nitrogen atom to a carbon atom is an ideal way to synthesize highly elaborate molecules in which nitrogen and oxygen functionalities are vicinally arranged. Recently, it has been disclosed that this intriguing transformation takes place in the presence of Lewis acidic metal catalysts. In this digest, we summarize recent findings on the catalytic [1,3]-oxygen rearrangement reactions, such as direct [1,3]-rearrangement of N-oxygenated enamines and anilines, as well as π-acidic metal-catalyzed cascade reaction involving the [1,3]-rearrangement process.