Direct N-glycosylation profiling of urine and prostatic fluid glycoproteins and extracellular vesicles

Expressed prostatic secretions (EPS), also called post digital rectal exam urines, are proximal fluids of the prostate that are widely used for diagnostic and prognostic assays for prostate cancer. These fluids contain an abundant number of glycoproteins and extracellular vesicles secreted by the prostate gland, and the ability to detect changes in their N-glycans composition as a reflection of disease state represents potential new biomarker candidates. Methods to characterize these N-glycan constituents directly from clinical samples in a timely manner and with minimal sample processing requirements are not currently available. In this report, an approach is described to directly profile the N-glycan constituents of EPS urine samples, prostatic fluids and urine using imaging mass spectrometry for detection. An amine reactive slide is used to immobilize glycoproteins from a few microliters of spotted samples, followed by peptide N-glycosidase digestion. Over 100 N-glycan compositions can be detected with this method, and it works with urine, urine EPS, prostatic fluids, and urine EPS-derived extracellular vesicles. A comparison of the N-glycans detected from the fluids with tissue N-glycans from prostate cancer tissues was done, indicating a subset of N-glycans present in fluids derived from the gland lumens. The developed N-glycan profiling is amenable to analysis of larger clinical cohorts and adaptable to other biofluids

Role of dynamical particle-vibration coupling in reconciliation of the d3/2d_{3/2} puzzle for spherical proton emitters

It has been observed that decay rate for proton emission from $d_{3/2}$ single particle state is systematically quenched compared with the prediction of a one dimensional potential model although the same model successfully accounts for measured decay rates from $s_{1/2}$ and $h_{11/2}$ states. We reconcile this discrepancy by solving coupled-channels equations, taking into account couplings between the proton motion and vibrational excitations of a daughter nucleus. We apply the formalism to proton emitting nuclei $^{160,161}$Re to show that there is a certain range of parameter set of the excitation energy and the dynamical deformation parameter for the quadrupole phonon excitation which reproduces simultaneously the experimental decay rates from the 2$d_{3/2}$, 3$s_{1/2}$ and 1$h_{11/2}$ states in these nuclei.Comment: RevTex, 12 pages, 4 eps figure

Hamiltonian structure of real Monge-Amp\`ere equations

The real homogeneous Monge-Amp\`{e}re equation in one space and one time dimensions admits infinitely many Hamiltonian operators and is completely integrable by Magri's theorem. This remarkable property holds in arbitrary number of dimensions as well, so that among all integrable nonlinear evolution equations the real homogeneous Monge-Amp\`{e}re equation is distinguished as one that retains its character as an integrable system in multi-dimensions. This property can be traced back to the appearance of arbitrary functions in the Lagrangian formulation of the real homogeneous Monge-Amp\`ere equation which is degenerate and requires use of Dirac's theory of constraints for its Hamiltonian formulation. As in the case of most completely integrable systems the constraints are second class and Dirac brackets directly yield the Hamiltonian operators. The simplest Hamiltonian operator results in the Kac-Moody algebra of vector fields and functions on the unit circle.Comment: published in J. Phys. A 29 (1996) 325

Relative spins and excitation energies of superdeformed bands in 190Hg: Further evidence for octupole vibration

An experiment using the Eurogam Phase II gamma-ray spectrometer confirms the existence of an excited superdeformed (SD) band in 190Hg and its very unusual decay into the lowest SD band over 3-4 transitions. The energies and dipole character of the transitions linking the two SD bands have been firmly established. Comparisons with RPA calculations indicate that the excited SD band can be interpreted as an octupole-vibrational structure.Comment: 12 pages, latex, 4 figures available via WWW at http://www.phy.anl.gov/bgo/bc/hg190_nucl_ex.htm

Advances in Quantitative Hepcidin Measurements by Time-of-Flight Mass Spectrometry

Assays for the detection of the iron regulatory hormone hepcidin in plasma or urine have not yet been widely available, whereas quantitative comparisons between hepcidin levels in these different matrices were thus far even impossible due to technical restrictions. To circumvent these limitations, we here describe several advances in time-of flight mass spectrometry (TOF MS), the most important of which concerned spiking of a synthetic hepcidin analogue as internal standard into serum and urine samples. This serves both as a control for experimental variation, such as recovery and matrix-dependent ionization and ion suppression, and at the same time allows value assignment to the measured hepcidin peak intensities. The assay improvements were clinically evaluated using samples from various patients groups and its relevance was further underscored by the significant correlation of serum hepcidin levels with serum iron indices in healthy individuals. Most importantly, this approach allowed kinetic studies as illustrated by the paired analyses of serum and urine samples, showing that more than 97% of the freely filtered serum hepcidin can be reabsorbed in the kidney. Thus, the here reported advances in TOF MS-based hepcidin measurements represent critical steps in the accurate quantification of hepcidin in various body fluids and pave the way for clinical studies on the kinetic behavior of hepcidin in both healthy and diseased states

Genome-Wide Screen of Three Herpesviruses for Protein Subcellular Localization and Alteration of PML Nuclear Bodies

Herpesviruses are large, ubiquitous DNA viruses with complex host interactions, yet many of the proteins encoded by these viruses have not been functionally characterized. As a first step in functional characterization, we determined the subcellular localization of 234 epitope-tagged proteins from herpes simplex virus, cytomegalovirus, and Epstein–Barr virus. Twenty-four of the 93 proteins with nuclear localization formed subnuclear structures. Twelve of these localized to the nucleolus, and five at least partially localized with promyelocytic leukemia (PML) bodies, which are known to suppress viral lytic infection. In addition, two proteins disrupted Cajal bodies, and 19 of the nuclear proteins significantly decreased the number of PML bodies per cell, including six that were shown to be SUMO-modified. These results have provided the first functional insights into over 120 previously unstudied proteins and suggest that herpesviruses employ multiple strategies for manipulating nuclear bodies that control key cellular processes

Relative K-stability for Kähler manifolds

We study the existence of extremal Kähler metrics on Kähler manifolds. After introducing a notion of relative K-stability for Kahler manifolds, we prove that Kähler manifolds admitting extremal Kähler metrics are relatively K-stable. Along the way, we prove a general Lp lower bound on the Calabi functional involving test configurations and their associated numerical invariants, answering a question of Donaldson. When the Kähler manifold is projective, our definition of relative K-stability is stronger than the usual definition given by Székelyhidi. In particular our result strengthens the known results in the projective case (even for constant scalar curvature Kähler metrics), and rules out a well known counterexample to the "naïve" version of the Yau-Tian-Donaldson conjecture in this setting