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The cytochrome P450s enzyme family metabolizes ∼80% of small molecule drugs. Variants in cytochrome P450s can substantially alter drug metabolism, leading to improper dosing and severe adverse drug reactions. Due to low sequence conservation, predicting variant effects across cytochrome P450s is challenging. Even closely related cytochrome P450s like CYP2C9 and CYP2C19, which share 92% amino acid sequence identity, display distinct phenotypic properties. Using variant abundance by massively parallel sequencing, we measured the steady-state protein abundance of 7,660 single amino acid variants in CYP2C19 expressed in cultured human cells. Our findings confirmed critical positions and structural features essential for cytochrome P450 function, and revealed how variants at conserved positions influence abundance. We jointly analyzed 4,670 variants whose abundance was measured in both CYP2C19 and CYP2C9, finding that the homologs have different variant abundances in substrate recognition sites within the hydrophobic core. We also measured the abundance of all single and some multiple wild type amino acid exchanges between CYP2C19 and CYP2C9. While most exchanges had no effect, substitutions in substrate recognition site 4 reduced abundance in CYP2C19. Double and triple mutants showed distinct interactions, highlighting a region that points to differing thermodynamic properties between the 2 homologs. These positions are known contributors to substrate specificity, suggesting an evolutionary tradeoff between stability and enzymatic function. Finally, we analyzed 368 previously unannotated human variants, finding that 43% had decreased abundance. By comparing variant effects between these homologs, we uncovered regions underlying their functional differences, advancing our understanding of this versatile family of enzymes., Competing Interests: Conflicts of interest The authors declare no conflict of interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Genetics Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Stem cells have lower facultative heterochromatin as defined by trimethylation of histone H3 lysine 27 (H3K27me3) compared to differentiated cells. However, the mechanisms underlying these differential H3K27me3 levels remain elusive. Because H3K27me3 levels are diluted two-fold in every round of replication and then restored through the rest of the cell cycle, we reasoned that the cell cycle length could be a key regulator of total H3K27me3 levels. Here, we propose that a fast cell cycle restricts H3K27me3 levels in stem cells. To test this model, we determined changes to H3K27me3 levels in mESCs globally and at specific loci upon G1 phase lengthening - accomplished by thymidine block or growth in the absence of serum (with the "2i medium"). H3K27me3 levels in mESC increase with G1 arrest when grown in serum and in 2i medium. Additionally, we observed via CUT&RUN and ChIP-seq that regions that gain H3K27me3 in G1 arrest and 2i media overlap, supporting our model of cell cycle length as a critical regulator of the stem cell epigenome and cellular identity. Furthermore, we demonstrate the inverse effect - that G1 shortening in differentiated cells results in a loss of H3K27me3 levels. Finally, in tumor cells with extreme H3K27me3 loss, lengthening of the G1 phase leads to H3K27me3 recovery despite the presence of the dominant negative, sub-stoichiometric H3.1K27M mutation. Our results indicate that G1 length is an essential determinant of H3K27me3 landscapes across diverse cell types.

Antisense oligonucleotide (ASO) therapy for neurological disease has been successful in clinical settings and its potential has generated hope for Alzheimer's disease (AD). We previously described that ablating SNCA encoding for α-synuclein (αSyn) in a mouse model of AD was beneficial. Here, we sought to demonstrate whether transient reduction of αSyn expression using ASO SNCA could be therapeutic in a mouse model of AD. The efficacy of the ASO SNCA was measured via immunocytochemistry, RT-qPCR and western blotting. To assess spatial learning and memory, ASO SNCA or PBS-injected APP and non-transgenic (NTG) mice, and separate groups of SNCA-null mice, were tested on the Barnes circular maze. Hippocampal slice electrophysiology and transcriptomic profiling were used to explore synaptic function and differential gene expression between groups. Reduction of SNCA transcripts alleviated cognitive deficits in male transgenic animals, but surprisingly, not in females. To determine the functional cause of this differential effect, we assessed memory function in SNCA-null mice. Learning and memory were intact in male mice but impaired in female animals, revealing that the role of αSyn on cognitive function is sex-specific. Transcriptional analyses identified a differentially expressed gene network centered around EGR1, a central modulator of learning and memory, in the hippocampi of SNCA-null mice. Thus, these novel results demonstrate that the function of αSyn on memory differs between male and female brains., (© 2022. The Author(s).)

Discerning an underlying structure in the array of magnetic resonance (MR) imaging sequences and acronyms available is a bewildering task. Attempts have been made to present standard taxonomies of MR imaging sequences, primarily on the basis of their underlying physics. Despite this, it is difficult to usefully incorporate given taxonomies into routine clinical knowledge. The links, differences, and similarities among sequences are multidimensional and too complex for tabular presentation on the printed page. The authors present an interactive taxonomy of MR imaging sequences. With this graphical interface, the user can explore the changing relationships among a wide range of sequence types as they are viewed from different perspectives and through different associations, working through a natural learning process.

Purpose: To determine retrospectively whether the anterior spinal artery (ASA) is visualized in the cervical region with contrast material-enhanced high-spatial-resolution three-dimensional magnetic resonance (MR) angiography of the extracranial carotid and vertebral arteries., Materials and Methods: The institutional research ethics committee approved this study and provided a waiver for informed consent. Data sets were evaluated in 50 consecutive patients referred for contrast-enhanced three-dimensional MR angiography of the carotid arteries (32 male and 18 female patients; age range, 15-80 years; mean age, 59 years). The ASA was defined as a linear area of high signal intensity that is seen anterior to the spinal cord in an arterial phase of enhancement and connects directly to a known arterial structure. If the linear area of high signal intensity was seen in the arterial phase but did not connect to a known arterial structure, it was considered a probable ASA. Venous enhancement was graded on a five-point scale (0-4) with grade 0 (no venous enhancement) or grade 1 (trace venous enhancement) considered to be in the arterial phase., Results: The ASA was identified with certainty in 37 of 50 patients. A vessel visualized anterior to the spinal cord, which probably represented the ASA, was seen in another 11 of 50 patients. In 29 of 50 patients the vessel was visualized only on the full-volume maximum intensity projection (MIP) image. In the remainder of cases the artery was identified on operator-defined subvolume MIP images. Continuity between the vessel and the vertebrobasilar arterial structures was identified in 35 of 50 patients. The vessel was seen as a continuous structure throughout its length in 34 patients and appeared discontinuous in 14. Radiculomedullary feeders were identified in 24 of 50 patients., Conclusion: The normal cervical ASA was visualized in 48 of 50 of subjects with contrast-enhanced high-spatial-resolution three-dimensional MR angiography.