Formation of mammalian erythrocytes: chromatin condensation and enucleation

In all vertebrates, the cell nucleus becomes highly condensed and transcriptionally inactive during the final stages of red cell biogenesis. Enucleation, the process by which the nucleus is extruded by budding off from the erythroblast, is unique to mammals. Enucleation has critical physiological and evolutionary significance in that it allows an elevation of hemoglobin levels in the blood and also gives red cells their flexible biconcave shape. Recent experiments reveal that enucleation involves multiple molecular and cellular pathways that include histone deacetylation, actin polymerization, cytokinesis, cell–matrix interactions, specific microRNAs and vesicle trafficking; many evolutionarily conserved proteins and genes have been recruited to participate in this uniquely mammalian process. In this review, we discuss recent advances in mammalian erythroblast chromatin condensation and enucleation, and conclude with our perspectives on future studies.National Institutes of Health (U.S.) (Grant P01 HL 32262)Amgen Inc. (Research Grant

Histone deacetylase 2 is required for chromatin condensation and subsequent enucleation of cultured mouse fetal erythroblasts

Background: During the final stages of differentiation of mammalian erythroid cells, the chromatin is condensed and enucleated. We previously reported that Rac GTPases and their downstream target, mammalian homolog of Drosophila diaphanous 2 (mDia2), are required for enucleation of in vitro cultured mouse fetal liver erythroblasts. However, it is not clear how chromatin condensation is achieved and whether it is required for enucleation. Design and Methods: Mouse fetal liver erythroblasts were purified from embryonic day 14.5 pregnant mice and cultured in erythropoietin-containing medium. Enucleation was determined by flow-cytometry based analysis after treatment with histone deacetylase inhibitors or infection with lentiviral short harirpin RNA. Results: We showed that histone deacetylases play critical roles in chromatin condensation and enucleation in cultured mouse fetal liver erythroblasts. Enzymatic inhibition of histone deacetylases by trichostatin A or valproic acid prior to the start of enucleation blocked chromatin condensation, contractile actin ring formation and enucleation. We further demonstrated that histone deacetylases 1, 2, 3 and 5 are highly expressed in mouse fetal erythroblasts. Short hairpin RNA down-regulation of histone deacetylase 2, but not of the other histone deacetylases, phenotypically mimicked the effect of trichostatin A or valproic acid treatment, causing significant inhibition of chromatin condensation and enucleation. Importantly, knock-down of histone deacetylase 2 did not affect erythroblast proliferation, differentiation, or apoptosis. Conclusions: These results identify histone deacetylase 2 as an important regulator, mediating chromatin condensation and enucleation in the final stages of mammalian erythropoiesis.National Institutes of Health (U.S.) (NIH grant P01 HL 32262)Amgen, Inc.National Institutes of Health (U.S.) (Pathway to Independence Award)Leukemia & Lymphoma Society of AmericaTemasek Life Sciences Laborator

Recurrent acquisition of cytosine methyltransferases into eukaryotic retrotransposons

Transposable elements are in a constant arms race with the silencing mechanisms of their host genomes. One silencing mechanism commonly used by many eukaryotes is dependent on cytosine methylation, a covalent modification of DNA deposited by C5 cytosine methyltransferases (DNMTs). Here, we report how two distantly related eukaryotic lineages, dinoflagellates and charophytes, have independently incorporated DNMTs into the coding regions of distinct retrotransposon classes. Concomitantly, we show that dinoflagellates of the genus Symbiodinium have evolved cytosine methylation patterns unlike any other eukaryote, with most of the genome methylated at CG dinucleotides. Finally, we demonstrate the ability of retrotransposon DNMTs to methylate CGs de novo, suggesting that retrotransposons could self-methylate retrotranscribed DNA. Together, this is an example of how retrotransposons incorporate host-derived genes involved in DNA methylation. In some cases, this event could have implications for the composition and regulation of the host epigenomic environment

Transiting Exoplanet Monitoring Project (TEMP). V. Transit Follow Up for HAT-P-9b, HAT-P-32b, and HAT-P-36b

© 2019. The American Astronomical Society. All rights reserved. During the past five years, 6, 7, and 26 transit observations were carried out for the HAT-P-9b, HAT-P-32b, and HAT-P-36b systems, respectively, through the Transiting Exoplanet Monitoring Project network. Combined with the published photometric data and radial-velocity measurements, our new photometry allows us to revisit the system parameters and search for additional close-in planetary companions in these hot Jupiter systems. We measure an updated R P /R ∗ = 0.1260 ±0.0011 for HAT-P-36 system in the R band, which is 4.5σ larger than the published i-band radius ratio of 0.1186 ±0.0012. We also perform a transit timing variation (TTV) analysis for each system. Because no significant TTVs were found, we place an upper mass limit on an additional planet for each system

Edoxaban: an update on the new oral direct factor Xa inhibitor.

Edoxaban is a once-daily oral anticoagulant that rapidly and selectively inhibits factor Xa in a concentration-dependent manner. This review describes the extensive clinical development program of edoxaban, including phase III studies in patients with non-valvular atrial fibrillation (NVAF) and symptomatic venous thromboembolism (VTE). The ENGAGE AF-TIMI 48 study (N = 21,105; mean CHADS2 score 2.8) compared edoxaban 60 mg once daily (high-dose regimen) and edoxaban 30 mg once daily (low-dose regimen) with dose-adjusted warfarin [international normalized ratio (INR) 2.0-3.0] and found that both regimens were non-inferior to warfarin in the prevention of stroke and systemic embolism in patients with NVAF. Both edoxaban regimens also provided significant reductions in the risk of hemorrhagic stroke, cardiovascular mortality, major bleeding and intracranial bleeding. The Hokusai-VTE study (N = 8,292) in patients with symptomatic VTE had a flexible treatment duration of 3-12 months and found that following initial heparin, edoxaban 60 mg once daily was non-inferior to dose-adjusted warfarin (INR 2.0-3.0) for the prevention of recurrent VTE, and also had a significantly lower risk of bleeding events. Both studies randomized patients at moderate-to-high risk of thromboembolic events and were further designed to simulate routine clinical practice as much as possible, with edoxaban dose reduction (halving dose) at randomisation or during the study if required, a frequently monitored and well-controlled warfarin group, a well-monitored transition period at study end and a flexible treatment duration in Hokusai-VTE. Given the phase III results obtained, once-daily edoxaban may soon be a key addition to the range of antithrombotic treatment options

A secular theory of coplanar, non-resonant planetary system

We present the secular theory of coplanar $N$-planet system, in the absence of mean motion resonances between the planets. This theory relies on the averaging of a perturbation to the two-body problem over the mean longitudes. We expand the perturbing Hamiltonian in Taylor series with respect to the ratios of semi-major axes which are considered as small parameters, without direct restrictions on the eccentricities. Next, we average out the resulting series term by term. This is possible thanks to a particular but in fact quite elementary choice of the integration variables. It makes it possible to avoid Fourier expansions of the perturbing Hamiltonian. We derive high order expansions of the averaged secular Hamiltonian (here, up to the order of 24) with respect to the semi-major axes ratio. The resulting secular theory is a generalization of the octupole theory. The analytical results are compared with the results of numerical (i.e., practically exact) averaging. We estimate the convergence radius of the derived expansions, and we propose a further improvement of the algorithm. As a particular application of the method, we consider the secular dynamics of three-planet coplanar system. We focus on stationary solutions in the HD 37124 planetary system.Comment: 14 pages, 4 figures, MATHEMATICA file expansion.m, accepted to Monthly Notices of the Royal Astronomical Society. (minor corrections of symbols

Transcription factors Foxo3a and Foxo1 couple the E3 ligase Cbl-b to the induction of Foxp3 expression in induced regulatory T cells

The transcription factor Foxp3 is essential for optimal regulatory T (T reg) cell development and function. Here, we show that CD4+ T cells from Cbl-b RING finger mutant knockin or Cbl-b–deficient mice show impaired TGF-β–induced Foxp3 expression. These T cells display augmented Foxo3a phosphorylation, but normal TGF-β signaling. Expression of Foxo3a rescues Foxp3 expression in Cbl-b–deficient T cells, and Foxo3a deficiency results in defective TGF-β–driven Foxp3 induction. A Foxo3a-binding motif is present in a proximal region of the Foxp3 promoter, and is required for Foxo3a association. Foxo1 exerts similar effects as Foxo3a on Foxp3 expression. This study reveals that Foxo factors promote transcription of the Foxp3 gene in induced T reg cells, and thus provides new mechanistic insight into Foxo-mediated T cell regulation

A Greatly Under-Appreciated Fundamental Principle of Physical Organic Chemistry

If a species does not have a finite lifetime in the reaction medium, it cannot be a mechanistic intermediate. This principle was first enunciated by Jencks, as the concept of an enforced mechanism. For instance, neither primary nor secondary carbocations have long enough lifetimes to exist in an aqueous medium, so SN1 reactions involving these substrates are not possible, and an SN2 mechanism is enforced. Only tertiary carbocations and those stabilized by resonance (benzyl cations, acylium ions) are stable enough to be reaction intermediates. More importantly, it is now known that neither H3O+ nor HO− exist as such in dilute aqueous solution. Several recent high-level calculations on large proton clusters are unable to localize the positive charge; it is found to be simply “on the cluster” as a whole. The lifetime of any ionized water species is exceedingly short, a few molecular vibrations at most; the best experimental study, using modern IR instrumentation, has the most probable hydrated proton structure as H13O6+, but only an estimated quarter of the protons are present even in this form at any given instant. Thanks to the Grotthuss mechanism of chain transfer along hydrogen bonds, in reality a proton or a hydroxide ion is simply instantly available anywhere it is needed for reaction. Important mechanistic consequences result. Any charged oxygen species (e.g., a tetrahedral intermediate) is also not going to exist long enough to be a reaction intermediate, unless the charge is stabilized in some way, usually by resonance. General acid catalysis is the rule in reactions in concentrated aqueous acids. The Grotthuss mechanism also means that reactions involving neutral water are favored; the solvent is already highly structured, so the entropy involved in bringing several solvent molecules to the reaction center is unimportant. Examples are given

Direct Imaging Explorations for Companions around Mid-Late M Stars from the Subaru/IRD Strategic Program

The Subaru telescope is currently performing a strategic program (SSP) using the high-precision near-infrared (NIR) spectrometer IRD to search for exoplanets around nearby mid/late-M~dwarfs via radial velocity (RV) monitoring. As part of the observing strategy for the exoplanet survey, signatures of massive companions such as RV trends are used to reduce the priority of those stars. However, this RV information remains useful for studying the stellar multiplicity of nearby M~dwarfs. To search for companions around such ``deprioritized" M~dwarfs, we observed 14 IRD-SSP targets using Keck/NIRC2 observations with pyramid wavefront sensing at NIR wavelengths, leading to high sensitivity to substellar-mass companions within a few arcseconds. We detected two new companions (LSPM~J1002+1459~B and LSPM~J2204+1505~B) and two new candidates that are likely companions (LSPM~J0825+6902~B and LSPM~J1645+0444~B) as well as one known companion. Including two known companions resolved by the IRD fiber injection module camera, we detected seven (four new) companions at projected separations between $\sim2-20$~au in total. A comparison of the colors with the spectral library suggests that LSPM~J2204+1505~B and LSPM~J0825+6902~B are located at the boundary between late-M and early-L spectral types. Our deep high-contrast imaging for targets where no bright companions were resolved did not reveal any additional companion candidates. The NIRC2 detection limits could constrain potential substellar-mass companions ($\sim10-75\ M_{\rm Jup}$) at 10~au or further. The failure with Keck/NIRC2 around the IRD-SSP stars having significant RV trends makes these objects promising targets for further RV monitoring or deeper imaging with JWST to search for smaller-mass companions below the NIRC2 detection limits.Comment: 16 pages, 8 figures, accepted for publication in A

Increased mitochondrial NADPH oxidase 4 (NOX4) expression in aging is a causative factor in aortic stiffening

Aging is characterized by increased aortic stiffness, an early, independent predictor and cause of cardiovascular disease. Oxidative stress from excess reactive oxygen species (ROS) production increases with age. Mitochondria and NADPH oxidases (NOXs) are two major sources of ROS in cardiovascular system. We showed previously that increased mitochondrial ROS levels over a lifetime induce aortic stiffening in a mouse oxidative stress model. Also, NADPH oxidase 4 (NOX4) expression and ROS levels increase with age in aortas, aortic vascular smooth muscle cells (VSMCs) and mitochondria, and are correlated with age-associated aortic stiffness in hypercholesterolemic mice. The present study investigated whether young mice (4 months-old) with increased mitochondrial NOX4 levels recapitulate vascular aging and age-associated aortic stiffness. We generated transgenic mice with low (Nox4TG605; 2.1-fold higher) and high (Nox4TG618; 4.9-fold higher) mitochondrial NOX4 expression. Young Nox4TG618 mice showed significant increase in aortic stiffness and decrease in phenylephrine-induced aortic contraction, but not Nox4TG605 mice. Increased mitochondrial oxidative stress increased intrinsic VSMC stiffness, induced aortic extracellular matrix remodeling and fibrosis, a leftward shift in stress-strain curves, decreased volume compliance and focal adhesion turnover in Nox4TG618 mice. Nox4TG618 VSMCs phenocopied other features of vascular aging such as increased DNA damage, increased premature and replicative senescence and apoptosis, increased proinflammatory protein expression and decreased respiration. Aortic stiffening in young Nox4TG618 mice was significantly blunted with mitochondrial-targeted catalase overexpression. This demonstration of the role of mitochondrial oxidative stress in aortic stiffness will galvanize search for new mitochondrial-targeted therapeutics for treatment of age-associated vascular dysfunction