Identification and analysis of the core biosynthetic machinery of tubulysin, a potent cytotoxin with potential anticancer activity

Myxobacteria are well known for their biosynthetic potential, especially for the production of cytotoxic compounds with potential anticancer activities. The tubulysins are currently in preclinical development. They are produced in very low quantities, and genetic manipulation of producing strains has never been accomplished. We report the development of a mariner-based transposon mutagenesis system for Angiococcus disciformis An d48. Extracts from a library of 1200 mutants were analyzed for the presence of tubulysin by a microscopic cell nucleus fragmentation bioassay. The transposition sites of four tubulysin-negative mutants were identified by vector recovery, which led to the identification and the sequencing of the corresponding core biosynthetic gene locus. Sequence analysis of more than 80,000 bp reveals an unusual multimodular hybrid polyketide synthase/peptide synthetase assembly line with a variety of unprecedented features.     

Millisecond pulsars with r-modes as steady gravitational radiators

Millisecond pulsars (MSPs) probably achieve their fast rotation by mass transfer from their companion stars in low-mass x-ray binaries (LMXBs). The lack of MSPs and LMXBs rotating near breakup has been attributed to the accretion torque being balanced, at fast rotation, by gravitational radiation, perhaps caused by an unstable oscillation mode. It has been argued that internal dissipation involving hyperons may cause LMXBs to evolve into a quasisteady state, with nearly constant rotation rate, temperature, and mode amplitude. We show that MSPs descending from these LMXBs spend a long time in a similar state, as extremely steady sources of gravitational waves and thermal x rays, while they spin down due to gravitational radiation and the standard magnetic torque. Observed MSP braking torques already place meaningful constraints on this scenario.     

Flow encoded NMR spectroscopy for quantification of metabolite flow in intact plants

An NMR flow quantification technique applicable to metabolite flow in plants is presented. It combines flow sensitive magnetization preparation with slice selective spectroscopy. Flow encoded NMR spectroscopy is described to quantify, for the first time, flow velocities of metabolites in plants non-invasively. Flow sensitivity is introduced by magnetization preparation based on a stimulated echo experiment, prior to slice selective spectroscopy. For flow quantification eight different flow-weighted spectra are collected. With this flow preparation very slow flow velocities down to 0.1mm/s can be detected and small amounts of flowing metabolites can be observed despite the large background signal of stationary and flowing water. Important sequence optimization steps include appropriate choice of experimental parameters used for flow encoding as well as complete balancing of eddy currents from the flow encoding gradients. The method was validated in phantom experiments and applied in vivo. Examples of quantitative flow measurements of water and metabolites in phantoms and plants are provided to demonstrate the reliability and the performance of flow encoded spectroscopy.     

NMR-microscopy with TrueFISP at 11.75T

The purpose of this paper is to demonstrate that a fully balanced gradient echo technique (TrueFISP) can be used for microscopic experiments at high static magnetic field strengths. TrueFISP experiments were successfully performed on homogeneous and inhomogeneous objects at 11.75T. High-resolution TrueFISP images were obtained from phantoms, plants, formalin-fixed samples, and from an isolated beating rat heart with an in-plane resolution of 78 micro m and a slice thickness of 500 micro m. The signal-to-noise ratio (SNR) gain of TrueFISP compared to conventional gradient echo or spin echo sequences will allow faster acquisition times or an improvement in spatial resolution for microscopic experiments.     

"Oscillating" metallocene catalysts: what stops the oscillation?

The 150 MHz (13)C NMR microstructural analysis of polypropylene samples produced with two representative "oscillating" metallocene catalysts of largely different steric hindrance, namely [(2-(3,5-di-tert-butyl-4-methoxyphenyl)indenyl)(2)ZrP](+) and [(2-phenylindenyl)(2)ZrP](+) (P = polymeryl), and the implications on the origin of the stereocontrol are presented and discussed in detail. The original mechanistic proposal of an "oscillation" between a rac-like (isotactic-selective) and a meso-like (nonstereoselective) conformation cannot explain the observed polymer configuration. The isotactic-stereoblock nature of the polymers obtained with the former catalyst proves unambiguously that the active cation "oscillates" between the two enantiomorphous rac-like conformations at an average frequency that, even at high propene concentration, is only slightly lower than that of monomer insertion. The less-hindered [(2-phenylindenyl)(2)ZrP](+) gives instead a largely stereoirregular polypropylene, which is the logical consequence of a faster ligand rotation; however, depending on the use conditions (in particular, on the nature of the cocatalyst and the polarity of the solvent), the polymerization products may also contain appreciable amounts of a fairly isotactic fraction. The peculiar microstructure of this fraction, with isotactic blocks of the same relative configuration spanned by very short atactic ones, rules out the possibility that the latter are due to an active species in meso-like conformation and points rather to a conformationally "locked" rac-like species with restricted ring mobility. The hypothesis of a stereorigidity induced by the proximity to a counteranion, which would play the role of the interannular bridge in the rac-bis(indenyl) ansa-metallocenes, was tested by computer modeling on a [rac-(2-phenylindenyl)(2)ZrMe(C(3)H(6))][B(C(6)F(5))(4)] ion couple and found viable.     

Carbon Monoxide Triplet Rydberg Series in the f Complex Region

Using a multistep state-selective excitation scheme, fluorescence-dip spectra of carbon monoxide have been recorded in the energy region between 106 100 and 112 200 cm(-1). Prominent groups of bands were tentatively identified as transitions into the 6f em leader10f (v=0) and 9psigma em leader11psigma (v=0) triplet Rydberg series. For the f complex states, molecular constants have been calculated. The triplet f levels show an unusually large rotational constant and are located at higher energies than their singlet counterparts. Copyright 2001 Academic Press.     

Differential algebras for quantum groups of type B, C, and D

We study higher order bicovariant differential calculi on the quantum groups O_q(N) and Sp _q (N). We show that the second antisymmetrizer exterior algebra _u is the quotient of the universal exterior algebra _u by the principal ideal generated by . Here denotes the unique up to scalars biinvariant 1-form. Moreover is central in _u and _u is an inner differential calculus. We show that the quadratic dual to the left-invariant algebra _{s L} is isomorphic to the reflection equation algebra. Let be an arbitrary left-covariant first order differential calculus. We show that the dimension of the space of left-invariant 2-forms in the universal exterior algebra equals the number of linearly independent quadratic-linear relations in the quantum tangent space.