76 research outputs found
An overview of the Michigan Positron Microscope Program
An overview of the Michigan Positron Microscope Program is presented with particular emphasis on the second generation microscope that is presently near completion. The design and intended applications of this microscope will be summarized.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87602/2/391_1.pd
Slow dynamics near glass transitions in thin polymer films
The -process (segmental motion) of thin polystyrene films supported
on glass substrate has been investigated in a wider frequency range from
10 Hz to 10 Hz using dielectric relaxation spectroscopy and thermal
expansion spectroscopy. The relaxation rate of the -process increases
with decreasing film thickness at a given temperature above the glass
transition. This increase in the relaxation rate with decreasing film thickness
is much more enhanced near the glass transition temperature. The glass
transition temperature determined as the temperature at which the relaxation
time of the -process becomes a macroscopic time scale shows a distinct
molecular weight dependence. It is also found that the Vogel temperature has
the thickness dependence, i.e., the Vogel temperature decreases with decreasing
film thickness. The expansion coefficient of the free volume is
extracted from the temperature dependence of the relaxation time within the
free volume theory. The fragility index is also evaluated as a function of
thickness. Both and are found to decrease with decreasing film
thickness.Comment: 9 pages, 7 figures, and 2 table
Casein kinase I Ī“/É phosphorylates topoisomerase IIĪ± at serine-1106 and modulates DNA cleavage activity
We previously reported that phosphorylation of topoisomerase (topo) IIĪ± at serine-1106 (Ser-1106) regulates enzyme activity and sensitivity to topo II-targeted drugs. In this study we demonstrate that phosphorylation of Ser-1106, which is flanked by acidic amino acids, is regulated in vivo by casein kinase (CK) IĪ“ and/or CKIÉ, but not by CKII. The CKI inhibitors, CKI-7 and IC261, reduced Ser-1106 phosphorylation and decreased formation of etoposide-stabilized topo IIāDNA cleavable complex. In contrast, the CKII inhibitor, 5,6-dichlorobenzimidazole riboside, did not affect etoposide-stabilized topo IIāDNA cleavable complex formation. Since, IC261 specifically targets the Ca2+-regulated isozymes, CKIĪ“ and CKIÉ, we examined the effect of down-regulating these enzymes on Ser-1106 phosphorylation. Down-regulation of these isozymes with targeted si-RNAs led to hypophosphorylation of the Ser-1106 containing peptide. However, si-RNA-mediated down-regulation of CKIIĪ± and Ī±ā² did not alter Ser-1106 phosphorylation. Furthermore, reduced phosphorylation of Ser-1106, observed in HRR25 (CKIĪ“/É homologous gene)-deleted Saccharomyces cerevisiae cells transformed with human topo IIĪ±, was enhanced following expression of human CKIÉ. Down-regulation of CKIĪ“ and CKIÉ also led to reduced formation of etoposide stabilized topo IIāDNA cleavable complex. These results provide strong support for an essential role of CKIĪ“/É in phosphorylating Ser-1106 in human topo IIĪ± and in regulating enzyme function
Quantitative proteomics reveals direct and indirect alterations in the histone code following methyltransferase knockdown
Histones are highly conserved proteins that organize cellular DNA. These proteins, especially their N-terminal domains, are adorned with many post-translational modifications (PTMs) such as lysine methylation, which are associated with active or repressed transcriptional states. The lysine methyltransferase G9a and its interaction partner Glp1 can mono- or dimethylate histone H3 on lysine (H3K9me1 or me2); possible cross-talk between these modifications and other PTMs on the same or other histone molecules is currently uncharacterized. In this study, we comprehensively analyze the effects of G9a/Glp1 knockdown on the most abundant histone modifications through both Bottom Up and Middle Down mass spectrometry-based proteomics. In addition to the expected decrease in H3K9me1/me2 we find that other degrees of methylation on K9 are affected by the reduction of G9a/Glp1 activity, particularly when K9 methylation occurs in combination with K14 acetylation. In line with this, an increase in K14 acetylation upon G9a knockdown was observed across all H3 variants (H3.1, H3.2 and H3.3), hinting at the potential existence of a binary switch between K9 methylation and K14 acetylation. Interestingly, we also detect changes in the abundance of other modifications (such as H3K79me2) in response to lowered levels of G9a/Glp1 suggesting histone PTM cross-talk amongst the H3 variants. In contrast, we find that G9a/Glp1 knockdown produces little effect on the levels of histone H4 PTMs, indicating low to no trans-histone PTM crosstalk. Lastly, we determined gene expression profiles of control and G9a/Glp1 knockdown cells, and we find that the G9a/Glp1 knockdown influences several genes, including DNA binding proteins and key factors in chromatin. Our results provide new insights into the intra- and inter- histone cross-regulation of histone K9 methylation and its potential downstream gene targets
Dramatic stiffening of ultrathin polymer films in the rubbery regime
Recently, we (P.A. O'Connell, G.B. McKenna, Science 307, 1760 (2005)) introduced a novel nano-bubble inflation method to measure the absolute creep compliance of nanometer thick polymer films. In that work it was shown that even at film thicknesses as small as 27.5nm the glass temperature was unchanged for poly(vinyl acetate) (PVAc). Perhaps more importantly, and the subject of the present work, was the observation that these ultrathin films show a dramatic stiffening in the rubbery plateau regime, i.e., the compliance was reduced by over two orders of magnitude compared to the bulk material. In the present work we substantiate the previous results in a study of the thickness dependence of the rubbery compliance of PVAc and polystyrene (PS) films for thicknesses from 13nm to 276nm. We show the substantial stiffening of the plateau region for both materials. Furthermore, the rubbery compliance (inverse of stiffness) scales with approximately the second power ( 1.8Ā±0.2) in the film thickness for both materials
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