Growth of protein crystals in hydrogels prevents osmotic shock

High-throughput protein X-ray crystallography offers a significant opportunity to facilitate drug discovery. The most reliable approach is to determine the three-dimensional structure of the protein-ligand complex by soaking the ligand in apo crystals. However, protein apo crystals produced by conventional crystallization in a solution are fatally damaged by osmotic shock during soaking. To overcome this difficulty, we present a novel technique for growing protein crystals in a high-concentration hydrogel that is completely gellified and exhibits high strength. This technique allowed us essentially to increase the mechanical stability of the crystals, preventing serious damage to the crystals caused by osmotic shock. Thus, this method may accelerate structure-based drug discoveries.     

Katorazone,a new yellow pigment with a 2-azaquinone-phenylhydrazone structure produced by Streptomyces sp. IFM 11299

An unusual alkaloid with a 2-azaquinone-phenylhydrazone structure, katorazone (1), and other metabolites were isolated from the ethyl acetate extract of Streptomyces sp. IFM 11299. The chemical structure of katorazone (1) was elucidated by 1D and 2D NMR analyses together with HR-ESI mass spectrometry. Katorazone (1) showed a synergistic effect in combination with TRAIL and decreased the viability of human gastric adenocarcinoma (AGS) cells.     

Crystal structure of stable protein CutA1 from psychrotrophic bacterium Shewanella sp. SIB1

CutA1 is widely found in bacteria, plants and animals, including humans. The functions of CutA1, however, have not been well clarified. It is known that CutA1s from Pyrococcus horikoshii, Thermus thermophilus and Oryza sativa unfold at temperatures remarkably higher than the growth temperatures of the host organisms. In this work the crystal structure of CutA1 from the psychrotrophic bacterium Shewanella sp. SIB1 (SIB1-CutA1) in a trimeric form was determined at 2.7?? resolution. This is the first crystal structure of a psychrotrophic CutA1. The overall structure of SIB1-CutA1 is similar to those of CutA1 from Homo sapiens, Escherichia coli, Pyrococcus horikoshii, Thermus thermophilus, Termotoga maritima, Oryza sativa and Rattus norvergicus. A peculiarity is observed in the β2 strand. The β2 strand is divided into two short β strands, β2a and β2b, in SIB1-CutA1. A thermal denaturation experiment revealed that SIB1-CutA1 does not unfold completely at 363?K at pH 7.0, although Shewanella sp. SIB1 cannot grow at temperatures exceeding 303?K. These results indicate that the trimeric structural motif of CutA1 is the critical factor in its unusually high stability and suggest that CutA1 needs to maintain its high stability in order to function, even in psychrotrophs.     

A Note on the Existence of Nonsmooth Nonconvex Optimization Problems

Sufficient conditions for the existence of a solution to an abstract optimization problem in Banach spaces are given, which do not rely on convexity, regularity properties or a straightforward coerciveness assumption. Applications to sparsity-constrained optimization and to problems from mechanics are provided.     

The Effect of pH on Emulsion Polymerization of Styrene in the Presence of an Amphoteric Emulsifier

The emulsion polymerization of styrene in the presence of an amphoteric emulsifier of the amino acid type, N-lauroyl-1-N, N-dimethyllysine (DMLL), has been studied at various pH values. The polymerization rate and the particle number concentration increased with increasing pH of the aqueous DMLL solution and these curves changed in slope at approximately pH 4 and pH 9. The physicochemical properties of the aqueous DMLL solution, such as CMC and the aggregation number, changed with the pH and similarly changed in slope at almost the same pH as the emulsion polymerization. These pH values were in good agreement with the pH at which the ionic form of the DMLL molecule changes. The number of latex particles formed changed in proportion to the number of DMLL micelles in the solution. The particle size and the molecular weight of the synthesized latex particles were also dependent on the physicochemical solution properties accompanying the change in the ionic forms of DMLL.     

Simultaneous determination of NG‐monomethyl‐l‐arginine,NG,NG‐dimethyl‐l‐arginine,NG,NG′‐dimethyl‐l‐arginine,and l‐arginine using monolithic silica disk‐packed spin columns and a monolithic silica column

We have developed and validated a high‐performance liquid chromatography method that uses monolithic silica disk‐packed spin columns and a monolithic silica column for the simultaneous determination of N^G‐monomethyl‐l ‐arginine, N^G,N^G‐dimethyl‐l ‐arginine, and N^G,N^G′‐dimethyl‐l ‐arginine in human plasma. For solid‐phase extraction, our method employs a centrifugal spin column packed with monolithic silica bonded to propyl benzenesulfonic acid as a cation exchanger. After pretreatment, the methylated arginines are converted to fluorescent derivatives with 4‐fluoro‐7‐nitro‐2,1,3‐benzoxadiazole, and then the derivatives are separated on a monolithic silica column. l ‐Arginine concentration was also determined in diluted samples. Standard calibration curves revealed that the assay was linear in the concentration range 0.2–1.0 μM for methylated arginines and 40–200 μM for l ‐arginine. Linear regression of the calibration curve yielded equations with correlation coefficients of 0.999 (r²). The sensitivity was satisfactory, with a limit of detection ranging from 3.75 to 9.0 fmol for all four compounds. The RSDs were 4.3–4.8% (intraday) and 3.0–6.8% (interday). When this method was applied to samples from six healthy donors, the detected concentrations of N^G‐monomethyl‐l ‐arginine, N^G,N^G‐dimethyl‐l ‐arginine, N^G,N^G′‐dimethyl‐l ‐arginine and l ‐arginine were 0.05 ± 0.01, 0.41 ± 0.07, 0.59 ± 0.11, and 83.8 ± 30.43 μM (n = 6), respectively.     

Variational approach to shape derivatives for a class of Bernoulli problems

The shape derivative of a functional related to a Bernoulli problem is derived without using the shape derivative of the state. The gradient information is combined with level set ideas in a steepest descent algorithm. Numerical examples show the feasibility of the approach.     

Homogenization and penalization of functional first-order PDE

In this paper we study the asymptotic behavior of viscosity solutions for a functional partial differential equation with a small parameter as the parameter tends to zero. We study simultaneous effects of homogenization and penalization in functional first-order PDE. We establish a convergence theorem in which the limit equation is identified with some first order PDE.