Shell structure at N=28 near the dripline: spectroscopy of 42^{42}Si, 43^{43}P and 44^{44}S

Measurements of the N=28 isotones 42Si, 43P and 44S using one- and two-proton knockout reactions from the radioactive beam nuclei 44S and 46Ar are reported. The knockout reaction cross sections for populating 42Si and 43P and a 184 keV gamma-ray observed in 43P establish that the d_{3/2} and s_{1/2} proton orbits are nearly degenerate in these nuclei and that there is a substantial Z=14 subshell closure separating these two orbits from the d_{5/2} orbit. The increase in the inclusive two-proton knockout cross section from 42Si to 44S demonstrates the importance of the availability of valence protons for determining the cross section. New calculations of the two-proton knockout reactions that include diffractive effects are presented. In addition, it is proposed that a search for the d_{5/2} proton strength in 43P via a higher statistics one-proton knockout experiment could help determine the size of the Z=14 closure.Comment: Phys. Rev. C, in pres

pp-sdsd shell gap reduction in neutron-rich systems and cross-shell excitations in 20^{20}O

Excited states in $^{20}$O were populated in the reaction $^{10}$Be($^{14}$C,$\alpha$) at Florida State University. Charged particles were detected with a particle telescope consisting of 4 annularly segmented Si surface barrier detectors and $\gamma$ radiation was detected with the FSU $\gamma$ detector array. Five new states were observed below 6 MeV from the $\alpha$-$\gamma$ and $\alpha$-$\gamma$-$\gamma$ coincidence data. Shell model calculations suggest that most of the newly observed states are core-excited 1p-1h excitations across the $N = Z = 8$ shell gap. Comparisons between experimental data and calculations for the neutron-rich O and F isotopes imply a steady reduction of the $p$-$sd$ shell gap as neutrons are added

iPSCORE: A Resource of 222 iPSC Lines Enabling Functional Characterization of Genetic Variation across a Variety of Cell Types.

Large-scale collections of induced pluripotent stem cells (iPSCs) could serve as powerful model systems for examining how genetic variation affects biology and disease. Here we describe the iPSCORE resource: a collection of systematically derived and characterized iPSC lines from 222 ethnically diverse individuals that allows for both familial and association-based genetic studies. iPSCORE lines are pluripotent with high genomic integrity (no or low numbers of somatic copy-number variants) as determined using high-throughput RNA-sequencing and genotyping arrays, respectively. Using iPSCs from a family of individuals, we show that iPSC-derived cardiomyocytes demonstrate gene expression patterns that cluster by genetic background, and can be used to examine variants associated with physiological and disease phenotypes. The iPSCORE collection contains representative individuals for risk and non-risk alleles for 95% of SNPs associated with human phenotypes through genome-wide association studies. Our study demonstrates the utility of iPSCORE for examining how genetic variants influence molecular and physiological traits in iPSCs and derived cell lines

Differential responses to lithium in hyperexcitable neurons from patients with bipolar disorder.

Bipolar disorder is a complex neuropsychiatric disorder that is characterized by intermittent episodes of mania and depression; without treatment, 15% of patients commit suicide. Hence, it has been ranked by the World Health Organization as a top disorder of morbidity and lost productivity. Previous neuropathological studies have revealed a series of alterations in the brains of patients with bipolar disorder or animal models, such as reduced glial cell number in the prefrontal cortex of patients, upregulated activities of the protein kinase A and C pathways and changes in neurotransmission. However, the roles and causation of these changes in bipolar disorder have been too complex to exactly determine the pathology of the disease. Furthermore, although some patients show remarkable improvement with lithium treatment for yet unknown reasons, others are refractory to lithium treatment. Therefore, developing an accurate and powerful biological model for bipolar disorder has been a challenge. The introduction of induced pluripotent stem-cell (iPSC) technology has provided a new approach. Here we have developed an iPSC model for human bipolar disorder and investigated the cellular phenotypes of hippocampal dentate gyrus-like neurons derived from iPSCs of patients with bipolar disorder. Guided by RNA sequencing expression profiling, we have detected mitochondrial abnormalities in young neurons from patients with bipolar disorder by using mitochondrial assays; in addition, using both patch-clamp recording and somatic Ca2+ imaging, we have observed hyperactive action-potential firing. This hyperexcitability phenotype of young neurons in bipolar disorder was selectively reversed by lithium treatment only in neurons derived from patients who also responded to lithium treatment. Therefore, hyperexcitability is one early endophenotype of bipolar disorder, and our model of iPSCs in this disease might be useful in developing new therapies and drugs aimed at its clinical treatment

Aberrant DNA Methylation in Human iPSCs Associates with MYC-Binding Motifs in a Clone-Specific Manner Independent of Genetics

Induced pluripotent stem cells (iPSCs) show variable methylation patterns between lines, some of which reflect aberrant differences relative to embryonic stem cells (ESCs). To examine whether this aberrant methylation results from genetic variation or non-genetic mechanisms, we generated human iPSCs from monozygotic twins to investigate how genetic background, clone, and passage number contribute. We found that aberrantly methylated CpGs are enriched in regulatory regions associated with MYC protein motifs and affect gene expression. We classified differentially methylated CpGs as being associated with genetic and/or non-genetic factors (clone and passage), and we found that aberrant methylation preferentially occurs at CpGs associated with clone-specific effects. We further found that clone-specific effects play a strong role in recurrent aberrant methylation at specific CpG sites across different studies. Our results argue that a non-genetic biological mechanism underlies aberrant methylation in iPSCs and that it is likely based on a probabilistic process involving MYC that takes place during or shortly after reprogramming

Substituted Imidazole of 5-Fluoro-2-[4-[(2-phenyl-1H-imidazol-5- yl)methyl]-1-piperazinyl]pyrimidine Inactivates Cytochrome P450 2D6 by Protein Adduction

detected by mass spectrometry indicate that the phenyl group on the imidazole ring of SCH 66712 is one site of oxidation by CYP2D6 and could lead to methylene quinone formation. Three other metabolites were also observed. For understanding the metabolic pathway that leads to CYP2D6 inactivation, metabolism studies with CYP2C9 and CYP2C19 were performed because neither of these enzymes is significantly inhibited by SCH 66712. The metabolites formed by CYP2C9 and CYP2C19 are the same as those seen with CYP2D6, although in different abundance. Modeling studies with CYP2D6 revealed potential roles of various active site residues in the oxidation of SCH 66712 and inactivation of CYP2D6 and showed that the phenyl group of SCH 66712 is positioned at 2.2 Å from the heme iron

Multiomic QTL mapping reveals phenotypic complexity of GWAS loci and prioritizes putative causal variants

Most GWAS loci are presumed to affect gene regulation; however, only ∼43% colocalize with expression quantitative trait loci (eQTLs). To address this colocalization gap, we map eQTLs, chromatin accessibility QTLs (caQTLs), and histone acetylation QTLs (haQTLs) using molecular samples from three early developmental-like tissues. Through colocalization, we annotate 10.4% (n = 540) of GWAS loci in 15 traits by QTL phenotype, temporal specificity, and complexity. We show that integration of chromatin QTLs results in a 2.3-fold higher annotation rate of GWAS loci because they capture distal GWAS loci missed by eQTLs, and that 5.4% (n = 13) of GWAS colocalizing eQTLs are early developmental specific. Finally, we utilize the iPSCORE multiomic QTLs to prioritize putative causal variants overlapping transcription factor motifs to elucidate the potential genetic underpinnings of 296 GWAS-QTL colocalizations

A Study on the Mechanism of Inhibition of Human P450 2D6 by Schering 66712 Using Metabolite and 3D Modeling Studies

vii, 19 p.The P450 enzymes are extremely important for human metabolism of both endogenous and exogenous compounds in the body, especially medicinal drugs. The inhibition of these enzymes by pharmaceuticals can lead to adverse drug reactions if drug levels build up to toxic levels. P450 2D6 is the second leading drug metabolizer of the P450 family and its inhibition by 5- fluoro-2-[4-[(2-phenyl-1H-imidazol-5-yl)methyl]-1-piperazinyl]pyrimidine (Schering 66712) was studied here via LC-MS and metabolite studies with P450 2D6, P450 3A4, P450 2C19, and P450 2C9. The P450s were assayed with SCH 66712 and ran on an LC-MS to look for potential metabolites. Only P450 2C9 showed an expected metabolite mass; the others did not show any hypothesized metabolites. Additionally, modeling studies with P450 2D6, P450 3A4 and P450 2C9 were conducted to compare substrate orientation and key amino acid moieties that may be involved in binding and inhibition