Absolute configuration of pterocidin,a potent inhibitor of tumor cell invasion from a marine-derived Streptomyces

During the course of screening natural products for the inhibitors of tumor cell invasion, pterocidin, a linear polyketide with a δ-lactone terminus, was rediscovered from a Streptomyces strain of a marine sediment-origin. A series of J-based configuration analyses and NOESY analysis, coupled with chemical derivatization and chiral anisotropy analysis, established the absolute stereochemistry of five asymmetric centers in this compound.     

Identification of Site-Specific Degradation in Bacterially Expressed Human Fibroblast Growth Factor 4 and Generation of an Aminoterminally Truncated,Stable Form

Fibroblast growth factor 4 (FGF4) is considered as a crucial gene for tumorigenesis in humans and the development of mammalian embryos. The secreted, mature form of human FGF4 is thought to be comprised of 175 amino acid residues (proline³² to leucine²⁰⁶, Pro³²-Leu²⁰⁶). Here, we found that bacterially expressed, 6× histidine (His)-tagged human FGF4 (Pro³²-Leu²⁰⁶) protein, referred to as HishFGF4, was unstable such as in phosphate-buffered saline. In these conditions, site-specific cleavage, including between Ser⁵⁴ and Leu⁵⁵, in HishFGF4 was identified. In order to generate stable human FGF4 derivatives, a 6× His-tagged human FGF4 (Leu⁵⁵-Leu²⁰⁶), termed HishFGF4L, was expressed in Escherichia coli. HishFGF4L could be purified from the supernatant of cell lysates by heparin column chromatography. In phosphate-buffered saline, HishFGF4L was considered as sufficiently stable. HishFGF4L exerted significant mitogenic activities in mouse embryonic fibroblast Balb/c 3T3 cells. In the presence of PD173074, an FGF receptor inhibitor, the growth-stimulating activity of HishFGF4L disappeared. Taken together, we suggest that HishFGF4L is capable of promoting cell growth via an authentic FGF signaling pathway. Our study provides a simple method for the production of a bioactive human FGF4 derivative in E. coli.     

Visualizing Hepatic Copper Release in Long–Evans Cinnamon Rats Using Single-Photon Emission Computed Tomography

The potential utility of an imaging agent for the detection of hepatic copper was investigated in a Wilson’s disease animal model. Solid-phase peptide synthesis was used to construct an imaging agent which consisted of a copper-binding moiety, taken from the prion protein, and a gamma ray-emitting indium radiolabel. Long–Evans Cinnamon (LEC) rats were used for the Wilson’s disease animal model. Our evaluation methodology consisted of administering the indium-labeled agent to both LEC and genetically healthy Long–Evans (LE) cohorts via a tail vein injection and following the pharmacokinetics with single-photon emission computed tomography (SPECT) over the course of an hour. The animals were then sacrificed and their livers necropsied. An additional control agent, lacking the copper-binding moiety, was used to gauge whether any change in the hepatic uptake might be caused by other physiological differences between the two animal models. LEC rats injected with the indium-labeled agent had roughly double the amount of hepatic radioactivity as compared to the healthy control animals. The control agent, without the copper-binding moiety, displayed a hepatic signal similar to that of the control LE animals. Additional intraperitoneal spiking with CuSO₄ in C57BL/6 mice also found that the pharmacokinetics of the indium-labeled, prion-based imaging agent is profoundly altered by exposure to in vivo pools of extracellular copper. The described SPECT application with this compound represented a significant improvement over a previous MRI application using the same base peptide sequence.     

Reliability tests for Weibull distribution with variational shape parameter based on sudden death lifetime data

In the traditional design of reliability tests for assuring the mean time to failure (MTTF) in Weibull distribution with shape and scale parameters, it has been assumed that the shape parameter in the acceptable and rejectable populations is the same fixed number. For the purpose of expanding applicability of the reliability testing, Hisada and Arizono have developed a reliability sampling scheme for assuring MTTF in the Weibull distribution under the conditions that shape parameters in the both populations do not necessarily coincide, and are specified as interval values, respectively. Then, their reliability test is designed using the complete lifetime data. In general, the reliability testing based on the complete lifetime data requires the long testing time. As a consequence, the testing cost becomes sometimes expensive. In this paper, for the purpose of an economical plan of the reliability test, we consider the sudden death procedure for assuring MTTF in Weibull distribution with variational shape parameter.     

Comparison of standard materials for the establishment of calibration curves in solid sampling graphite furnace atomic absorption spectrometry. Direct determination of copper in powdered biological samples

Summary The selection of standard materials for the establishment of calibration curves is one of the most important problems in the direct analysis of biological samples by GFAAS. Three kinds of standard materials, NBS tomato leaves, coprecipitates with magnesium oxinate and Ni/DMG/PAN were investigated. It was found that it is possible to use both the NBS-SRM and the synthetic reference material prepared by coprecipitation with magnesium oxinate as standard materials for the direct determination of copper in several biological samples issued from NBS and NIES. However, too low results are obtained when the synthetic reference material prepared by coprecipitation with Ni/DMG/PAN is used. In order to clarify this, the effects of nickel and magnesium were examined and the role of magnesium was discussed.     

Numerical examination of the extended phase‐space volume‐preserving integrator by the Nosé‐Hoover molecular dynamics equations

This article illustrates practical applications to molecular dynamics simulations of the recently developed numerical integrators [Phys Rev E 2006, 73, 026703] for ordinary differential equations. This method consists of extending any set of ordinary differential equations in order to define a time invariant function, and then use the techniques of divergence‐free solvable decomposition and symmetric composition to obtain volume‐preserving integrators in the extended phase space. Here, we have developed the technique by constructing multiple extended‐variable formalism in order to enhance the handling in actual simulation, and by constituting higher order integrators to obtain further accuracies. Using these integrators, we perform constant temperature molecular dynamics simulations of liquid water, liquid argon and peptide in liquid water droplet. The temperature control is obtained through an extended version of the Nosé‐Hoover equations. Analyzing the effects of the simulation conditions including time step length, initial values, boundary conditions, and equation parameters, we investigate local accuracy, global accuracy, computational cost, and sensitivity along with the sampling validity. According to the results of these simulations, we show that the volume‐preserving integrators developed by the current method are more effective than traditional integrators that lack the volume‐preserving property. © 2008 Wiley Periodicals, Inc. J Comput Chem, 2009     

Synthesis and structural revision of marine eicosanoid agardhilactone

Synthesis of the marine eicosanoid agardhilactone has been achieved. The relative and absolute configuration of agardhilactone was successfully determined.