For pharmacokinetic modeling of tissue physiology, there is great interest in measuring the arterial input function (AIF) from dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) using paramagnetic contrast agents. Due to relaxation effects, the measured signal is a nonlinear function of the injected contrast agent concentration and depends on sequence parameters, system calibration, and time-of-flight effects, making it difficult to accurately measure the AIF during the first pass. Paramagnetic contrast agents also affect susceptibility and modify the magnetic field in proportion to their concentration. This information is contained in the MR signal phase which is discarded in a typical image reconstruction. However, quantifying AIF through contrast agent susceptibility induced phase changes is made difficult by the fact that the induced magnetic field is nonlocal and depends upon the contrast agent spatial distribution and thus on organ and vessel shapes. In this article, the contrast agent susceptibility was quantified through inversion of magnetic field shifts using a piece-wise constant model. Its feasibility is demonstrated by a determination of the AIF from the susceptibility-induced field changes of an intravenous bolus. After in vitro validation, a time-resolved two-dimensional (2D) gradient echo scan, triggered to diastole, was performed in vivo on the aortic arch during a bolus injection of 0.1 mmol/kg Gd-DTPA. An approximate geometrical model of the aortic arch constructed from the magnitude images was used to calculate the spatial variation of the field associated with the bolus. In 14 subjects, Gd concentration curves were measured dynamically (one measurement per heart beat) and indirectly validated by independent 2D cine phase contrast flow rate measurements. Flow rate measurements using indicator conservation with this novel quantitative susceptibility imaging technique were found to be in good agreement with those obtained from the cine phase contrast measurements in all subjects. Contrary to techniques that rely on intensity, the accuracy of this signal phase based method is insensitive to factors influencing signal intensity such as flip angle, coil sensitivity, relaxation changes, and time-of-flight effects extending the range of pulse sequences and contrast doses for which quantitative DCE-MRI can be applied.

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Genes adjacent to telomeres are subject to transcriptional repression mediated by an integrated set of chromatin modifying and remodeling factors. The telomeres of Saccharomyces cerevisiae have served as a model for dissecting the function of diverse chromatin proteins in gene silencing, and their study has revealed overlapping roles for many chromatin proteins in either promoting or antagonizing gene repression. The H3K4 methyltransferase Set1, which is commonly linked to transcriptional activation, has been implicated in telomere silencing. Set5 is an H4 K5, K8, and K12 methyltransferase that functions with Set1 to promote repression at telomeres. Here, we analyzed the combined role for Set1 and Set5 in gene expression control at native yeast telomeres. Our data reveal that Set1 and Set5 promote a Sir proteinindependent mechanism of repression that may primarily rely on regulation of H4K5ac and H4K8ac at telomeric regions. Furthermore, cells lacking both Set1 and Set5 have highly correlated transcriptomes to mutants in telomere maintenance pathways and display defects in telomere stability, linking their roles in silencing to protection of telomeres. Our data therefore provide insight into and clarify potential mechanisms by which Set1 contributes to telomere silencing and shed light on the function of Set5 at telomeres.

The conserved yeast histone methyltransferase Set1 targets H3 lysine 4 (H3K4) for mono, di, and trimethylation and is linked to active transcription due to the euchromatic distribution of these methyl marks and the recruitment of Set1 during transcription. However, loss of Set1 results in increased expression of multiple classes of genes, including genes adjacent to telomeres and middle sporulation genes, which are repressed under normal growth conditions because they function in meiotic progression and spore formation. The mechanisms underlying Set1-mediated gene repression are varied, and still unclear in some cases, although repression has been linked to both direct and indirect action of Set1, associated with noncoding transcription, and is often dependent on the H3K4me2 mark. We show that Set1, and particularly the H3K4me2 mark, are implicated in repression of a subset of middle sporulation genes during vegetative growth. In the absence of Set1, there is loss of the DNA-binding transcriptional regulator Sum1 and the associated histone deacetylase Hst1 from chromatin in a locus-specific manner. This is linked to increased H4K5ac at these loci and aberrant middle gene expression. These data indicate that, in addition to DNA sequence, histone modification status also contributes to proper localization of Sum1. Our results also show that the role for Set1 in middle gene expression control diverges as cells receive signals to undergo meiosis. Overall, this work dissects an unexplored role for Set1 in gene-specific repression, and provides important insights into a new mechanism associated with the control of gene expression linked to meiotic differentiation.

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