Separation of proteins by hydrophobic interaction chromatography at low salt concentration

We investigated protein separation by hydrophobic interaction chromatography (HIC) at low salt concentration on the supports of various hydrophobicities. Hydrophobic proteins could be successfully separated with more than 90% recovery by gradient elution of ammonium sulfate from 0.3-0.5 M to 0 in 50 mM phosphate buffer (pH 6.8) by using supports whose hydrophobicities were properly adjusted individually for each protein. Satisfactory results were also obtained by isocratic elution without ammonium sulfate and gradient elution of ethanol from 0 to 10%. HIC at low salt concentration was compatible with other modes of liquid chromatography like ion-exchange chromatography. On the other hand, it was not successful to separate hydrophilic proteins at low salt concentration. Recoveries of hydrophilic proteins decreased before they were retained enough as support hydrophobicity increased. Therefore, it is inevitable to use a higher concentration of salt, e.g., 1-2 M ammonium sulfate, on hydrophilic or moderately hydrophobic support in order to retain hydrophilic proteins without decrease in recovery.     

High-throughput evaluation of quiescent hematopoietic progenitor cells using a micro-multiwell plate

Conventional assays for hematopoietic progenitor cells (HPCs) require long-term culturing, a labor-intensive procedure, and technique proficiency. We aimed to develop a high-throughput method to determine frequency of quiescent primitive HPCs by a combination of the micro-multiwell plate and 5-fluorouracil (5-FU) treatment. The micro-multiwell plate was made of a silicone sheet with a 6 × 6 array of 1-mm diameter holes and a glass substrate. To enrich primitive HPCs in a CD34 population, CD34 cells and stromal cells were applied to micro-multiwells and cultured in the presence of 5-FU for 2 days. The quiescent primitive HPCs that survived after 5-FU treatment were then expanded with cytokines in the absence of 5-FU for a further 10 days. After culturing, cells were immunostained and the number of primitive HPCs in inoculated CD34 cells was estimated from fluorescent intensity for each well under a stereoscopic fluorescent microscope. The frequencies of primitive HPCs correlated well with frequencies of cobblestone area-forming cells for two CD34 cell lots. Our method allows high-throughput screening for primitive HPCs in CD34 cells. Figure Representative image of a micro-multiwell plate fordetecting primitive hematopoietic stem cells     

Single Crystals of Mixed-Cation Copper-Based Perovskite with Trimodal Bandgap Behavior

Two-dimensional (2D) hybrid perovskites with novel functionalities and structural diversity are a perfect platform for emerging optoelectronic devices such as photodetectors, light-emitting diodes, and solar cells. Here, we demonstrate that excess concentration of Cesium bromide (CsBr) is key to the formation of easily exfoliated 2D Cs₂Cu(Cl/Br)₄ perovskite crystal. Furthermore, by employing this trick to 2D perovskite MA₂Cu(Cl/Br)₄ (MA=methylammonium), we achieve a phase-pure easily exfoliated 2D mixed-cation (MA/Cs)₂Cu(Cl/Br)₄ perovskite crystal, which exhibits reduced bandgap (1.53 eV) with ferromagnetic behavior and photovoltaic property. The resultant mixed-cation structured device reveals enhanced efficiency compared to all MA and all Cs counterparts. These findings demonstrate the importance of cation-engineering in developing innovative materials with novel properties.     

Twisted mass quarks and the phase structure of lattice QCD

The phase structure of zero temperature twisted mass lattice QCD is investigated. We find strong metastabilities in the plaquette observable in correspondence of which the untwisted quark mass assumes positive or negative values. We provide interpretations of this phenomenon in terms of chiral symmetry breaking and the effective potential model of Sharpe and Singleton.Received: 24 August 2004, Revised: 29 October 2004, Published online: 25 January 2005     

Symmetric submanifolds associated with irreducible symmetric R-spaces

We construct examples of symmetric submanifolds in Riemannian symmetric spaces of noncompact type and obtain the classification of symmetric submanifolds in irreducible Riemannian symmetric spaces of noncompact type and rank greater than one. This finishes the classification problem of symmetric submanifolds in Riemannian symmetric spaces completely.     

Dynamical fluctuation spectra of two-dimensional classical electron liquids in magnetic fields

Dynamical fluctuation spectra of a two-dimensional classical system of electrons in a magnetic field are obtained by numerical experiments in the domain of a strongly coupled liquid. Excitations in these systems have dispersion relations similar to those in the Wigner lattice of the same number density.     

Gel chromatographic investigation of the substitution reaction between diphosphonate and hypophosphate

Gel chromatography was used to characterize the substitution reaction between diphosphonate and hypophosphate. In addition to a well-known trimeric oxo acid, ³P-O-⁴P-⁴P, a hitherto unknown tetrameric oxo acid, ³P-O-⁴P-⁴P-O-³P, was detected. The stability of the tetramer in aqueous solution was examined.     

Synthesis of indolizines and pyrrolo[2,1-b]azoles from 2-pyridylmagnesium bromide and 2-lithiated azoles using tris(alkylthio)-cyclopropenyl cations as a three-carbon building block

A convenient method for the preparation of indolizines 3 and 4 and pyrrolo[2,1-b]azoles 7a-e from 2-pyridylmagnesium bromide and 2-lithiated azoles 6a-e , respectively, was developed by using tris(tert-butylthio)- and tris(isopropylthio)cyclopropenyl cations 1 and 2 as a three-carbon building block.     

Selective determination of total normal microcystin by colorimetry, LC/UV detection and/or LC/MS

Microcystins, hepatotoxic cyclic heptapeptides, are produced by freshwater cyanobacteria, and are classified four groups according to the amino acid structure at unit 7. Normal microcystins contain N-methyldehydroalanine (Mdha) or dehydroalanine (Dha) at unit 7, and command the great part of all microcystins. As unusual microcystin classes, [Dhb⁷]microcystins, [ - and -Ala⁷, or N-MeAla⁷]microcystins and [ -Ser⁷]microcystins have been found.

On tumor initiation and/or promotion activities of microcystins, the tumor promotion activity of normal microcystins has been found, but cancer-related activities of microcystins belonging in the other classes have not been clear.

To determine normal microcystins as hepatotoxic tumor promoters, a selective determination method was developed. Only Mdha or Dha in normal microcystins was reacted with glutathione (GSH). The GSH-normal microcystins conjugates were reacted with trinitrobenzene sulfonate (TNBS). The TNB–GSH-normal microcystin conjugate can be determined as the total normal microcystin by colorimetry. After methanolysis of the conjugate, dimethyl TNB–glutamate from the conjugate was determined by liquid chromatography/ultraviolet detection (LC/UV) and/or liquid chromatography/mass spectrometry (LC/MS). The detection limits of the total normal microcystin by colorimetry, LC/UV and/or LC/MS were 1 μg, 10 and 0.1 ng, respectively.