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Objectives To investigate key factors that may contribute to the variability of rituximab-mediated peripheral and renal B cell depletion (BCD) in SLE. Methods We analysed: (i) CD19+ B cell counts in patients with SLE before and 1, 2, 3 and 6 months after treatment with rituximab, comparing them with RA patients; (ii) the presence of B cells in renal biopsies after rituximab therapy; (iii) whether the duration of BCD correlated with patient demographics and B cell expression of CD20 and FcγRIIb; and (iv) the effect of B cell activation factor (BAFF) on the efficiency of rituximab and obinutuzumab at inducing BCD in whole blood assays, in vitro. Results In SLE (n = 71), the duration of BCD was shorter compared with RA (n = 27). B cells were detectable in renal biopsy samples (n = 6) after treatment with rituximab in all patients with poor response while peripheral blood B cells remained low or undetectable in the same patients. There were no significant relationships between peripheral BCD and patient age, disease duration, serum C3 levels or the level of expression of B cell surface proteins CD20 and FcγRIIb. Obinutuzumab was more efficient than rituximab at inducing BCD in whole blood assays, regardless of excess BAFF. Conclusions BCD in SLE is less efficient than in RA. Renal B cell presence following rituximab treatment was associated with poor outcomes. No significant relationships between any measured B cell related, clinical or laboratory parameters and the efficiency of BCD by rituximab was found. Obinutuzumab was superior to rituximab at inducing BCD.

We prove that a "random" free group outer automorphism is an ageometric fully irreducible outer automorphism whose ideal Whitehead graph is a union of triangles. In particular, we show that its attracting (and repelling) tree is a nongeometric $\mathbb R$-tree all of whose branch points are trivalent, Comment: 27 pages comments are welcome!

We study properties of generic elements of groups of isometries of hyperbolic spaces. Under general combinatorial conditions, we prove that loxodromic elements are generic (i.e., they have full density with respect to counting in balls for the word metric in the Cayley graph) and translation length grows linearly. We provide applications to a large class of relatively hyperbolic groups and graph products, including all right-angled Artin groups and right-angled Coxeter groups.

Tet enzymes (Tet1/2/3) oxidize 5-methylcytosine to promote DNA demethylation and partner with chromatin modifiers to regulate gene expression. Tet1 is highly expressed in embryonic stem cells (ESCs), but its enzymatic and non-enzymatic roles in gene regulation are not dissected. We have generated Tet1 catalytically inactive (Tet1m/m) and knockout (Tet1−/−) ESCs and mice to study these functions. Loss of Tet1, but not loss of its catalytic activity, caused aberrant upregulation of bivalent (H3K4me3+; H3K27me3+) developmental genes, leading to defects in differentiation. Wild-type and catalytic-mutant Tet1 occupied similar genomic loci which overlapped with H3K27 tri-methyltransferase PRC2 and the deacetylase complex Sin3a at promoters of bivalent genes and with the helicase Chd4 at active genes. Loss of Tet1, but not loss of its catalytic activity, impaired enrichment of PRC2 and Sin3a at bivalent promoters leading to reduced H3K27 trimethylation and deacetylation, respectively, in absence of any changes in DNA methylation. Tet1−/−, but not Tet1m/m, embryos expressed higher levels of Gata6 and were developmentally delayed. Thus, the critical functions of Tet1 in ESCs and early development are mediated through its non-catalytic roles in regulating H3K27 modifications to silence developmental genes, and are more important than its catalytic functions in DNA demethylation.

Mosaic evolution refers to the pattern whereby different organismal traits exhibit differential rates of evolution typically due to reduced levels of trait covariation through deep time (i.e., modularity). These differences in rates can be attributed to variation in responses to selective pressures between individual traits. Differential responses to selective pressures also have the potential to facilitate functional specialization, allowing certain traits to track environmental stimuli more closely than others. The teleost skull is a multifunctional structure comprising a complex network of bones and thus an excellent system for which to study mosaic evolution. Here we construct an ultrametric phylogeny for a clade of Neotropical electric fishes (Apteronotidae: Gymnotiformes) and use three-dimensional geometric morphometrics to investigate patterns of mosaic evolution in the skull and jaws. We find strong support for a developmental, three-module hypothesis that consists of the face, braincase, and mandible, and we find that the mandible has evolved four times faster than its neighboring modules. We hypothesize that the functional specialization of the mandible in this group of fishes has allowed it to outpace the face and braincase and evolve in a more decoupled manner. We also hypothesize that this pattern of mosaicism may be widespread across other clades of teleost fishes.

MDMX is overexpressed in the vast majority of patients with acute myeloid leukemia (AML). We report that MDMX overexpression increases preleukemic stem cell (pre-LSC) number and competitive advantage. Utilizing five newly generated murine models, we found that MDMX overexpression triggers progression of multiple chronic/asymptomatic preleukemic conditions to overt AML. Transcriptomic and proteomic studies revealed that MDMX overexpression exerts this function, unexpectedly, through activation of Wnt/β-Catenin signaling in pre-LSCs. Mechanistically, MDMX binds CK1α and leads to accumulation of β-Catenin in a p53-independent manner. Wnt/β-Catenin inhibitors reverse MDMX-induced pre-LSC properties, and synergize with MDMX-p53 inhibitors. Wnt/β-Catenin signaling correlates with MDMX expression in patients with preleukemic myelodysplastic syndromes and is associated with increased risk of progression to AML. Our work identifies MDMX overexpression as a pervasive preleukemic-to-AML transition mechanism in different genetically driven disease subtypes, and reveals Wnt/β-Catenin as a non-canonical MDMX-driven pathway with therapeutic potential for progression prevention and cancer interception.

For certain pseudo-Anosov flows $\phi$ on closed $3$-manifolds, unpublished work of Agol--Gu\'eritaud produces a veering triangulation $\tau$ on the manifold $M$ obtained by deleting $\phi$'s singular orbits. We show that $\tau$ can be realized in $M$ so that its 2-skeleton is positively transverse to $\phi$, and that the combinatorially defined flow graph $\Phi$ embedded in $M$ uniformly codes $\phi$'s orbits in a precise sense. Together with these facts we use a modified version of the veering polynomial, previously introduced by the authors, to compute the growth rates of $\phi$'s closed orbits after cutting $M$ along certain transverse surfaces, thereby generalizing work of McMullen in the fibered setting. These results are new even in the case where the transverse surface represents a class in the boundary of a fibered cone of $M$. Our work can be used to study the flow $\phi$ on the original closed manifold. Applications include counting growth rates of closed orbits after cutting along closed transverse surfaces, defining a continuous, convex entropy function on the `positive' cone in $H^1$ of the cut-open manifold, and answering a question of Leininger about the closure of the set of all stretch factors arising as monodromies within a single fibered cone of a $3$-manifold. This last application connects to the study of endperiodic automorphisms of infinite-type surfaces and the growth rates of their periodic points., Comment: 75 pages, 32 figures. Final version to appear in Ergodic Theory and Dynamical Systems

The transformed state in acute leukemia requires gene regulatory programs involving transcription factors and chromatin modulators. Here, we uncover an IRF8-MEF2D transcriptional circuit as an acute myeloid leukemia (AML)-biased dependency. We discover and characterize the mechanism by which the chromatin “reader” ZMYND8 directly activates IRF8 in parallel with the MYC proto-oncogene through their lineage-specific enhancers. ZMYND8 is essential for AML proliferation in vitro and in vivo and associates with MYC and IRF8 enhancer elements that we define in cell lines and in patient samples. ZMYND8 occupancy at IRF8 and MYC enhancers requires BRD4, a transcription coactivator also necessary for AML proliferation. We show that ZMYND8 binds to the ET domain of BRD4 via its chromatin reader cassette, which in turn is required for proper chromatin occupancy and maintenance of leukemic growth in vivo. Our results rationalize ZMYND8 as a potential therapeutic target for modulating essential transcriptional programs in AML.

We prove that any mapping torus of a pseudo-Anosov mapping class with bounded normalized Weil-Petersson translation length contains a finite set of transverse and level closed curves, and drilling out this set of curves results in one of a finite number of cusped hyperbolic 3-manifolds. The number of manifolds in the finite list depends only on the bound for normalized translation length. We also prove a complementary result that explains the necessity of removing level curves by producing new estimates for the Weil-Petersson translation length of compositions of pseudo-Anosov mapping classes and arbitrary powers of a Dehn twist., v2. Added references. v1. 49 pages, 9 figures

IL1RAP is an emerging target for AML therapy. Studying its cell-intrinsic function revealed that IL1RAP interacts with and amplifies signaling through c-KIT and FLT3 in AML cells. This novel promiscuous role of IL1RAP in AML has implications for therapeutic targeting., The surface molecule interleukin-1 receptor accessory protein (IL1RAP) is consistently overexpressed across multiple genetic subtypes of acute myeloid leukemia (AML) and other myeloid malignancies, including at the stem cell level, and is emerging as a novel therapeutic target. However, the cell-intrinsic functions of IL1RAP in AML cells are largely unknown. Here, we show that targeting of IL1RAP via RNA interference, genetic deletion, or antibodies inhibits AML pathogenesis in vitro and in vivo, without perturbing healthy hematopoietic function or viability. Furthermore, we found that the role of IL1RAP is not restricted to the IL-1 receptor pathway, but that IL1RAP physically interacts with and mediates signaling and pro-proliferative effects through FLT3 and c-KIT, two receptor tyrosine kinases with known key roles in AML pathogenesis. Our study provides a new mechanistic basis for the efficacy of IL1RAP targeting in AML and reveals a novel role for this protein in the pathogenesis of the disease., Graphical Abstract

We study the connections between subsurface projections in curve and arc complexes in fibered 3-manifolds and Agol's veering triangulation. The main theme is that large-distance subsurfaces in fibers are associated to large simplicial regions in the veering triangulation, and this correspondence holds uniformly for all fibers in a given fibered face of the Thurston norm., Referee's comments incorporated. To appear in GAFA

We prove that finitely generated purely loxodromic subgroups of a right-angled Artin group A(Γ) fulfill equivalent conditions that parallel characterizations of convex cocompactness in mapping class groups Mod(S). In particular, such subgroups are quasiconvex in A(Γ). In addition, we identify a milder condition for a finitely generated subgroup of A(Γ) that guarantees it is free, undistorted, and retains finite generation when intersected with A(Λ) for subgraphs Λ of Γ. These results have applications to both the study of convex cocompactness in Mod(S) and the way in which certain groups can embed in right-angled Artin groups.

We introduce the co-surface graph $\mathcal{CS}$ of a finitely generated free group $\mathbb{F}$ and use it to study the geometry of hyperbolic group extensions of $\mathbb{F}$. Among other things, we show that the Gromov boundary of the co-surface graph is equivariantly homeomorphic to the space of free arational $\mathbb{F}$-trees and use this to prove that a finitely generated subgroup of $\mathrm{Out}(\mathbb{F})$ quasi-isometrically embeds into the co-surface graph if and only if it is purely atoroidal and quasi-isometrically embeds into the free factor complex. This answers a question of I. Kapovich. Our earlier work [Hyperbolic extensions of free groups, arXiv:1406.2567] shows that every such group gives rise to a hyperbolic extension of $\mathbb{F}$, and here we prove a converse to this result that characterizes the hyperbolic extensions of $\mathbb{F}$ arising in this manner. As an application of our techniques, we additionally obtain a Scott--Swarup type theorem for this class of extensions., 33 pages. Minor changes and other updates to incorporate referee comments. Final version; accepted for publication in the Journal of Topology

The p53 inhibitor MDMX is overexpressed in the vast majority of patients with AML; however, a causative role of MDMX in leukemogenesis has not yet been investigated. Mdmx transgenic mice (Mdmx-Tg) displayed increased number, proliferation and competitive advantage of hematopoietic stem cells compared to WT littermates. Crossing Mdmx-Tg with preleukemic mouse models representing frequent alterations in human AML, we generated three distinct compound models. We found that Mdmx overexpression triggered progression of each of the different chronic/asymptomatic preleukemic conditions to overt AML, and thereby markedly shortened survival. RNA sequencing revealed that Mdmx overexpression exerted this function through activation of Wnt/b-Catenin signaling in pre-LSC. Mechanistically, we found that MDMX bound CK1a, a canonical degrader of b-Catenin, and led to accumulation and nuclear translocation of b-Catenin. WNT/b-Catenin inhibitors or exogenous overexpression of CK1a were both effective in reversing Mdmx-induced pre-LSC properties, and showed further synergism in combination with MDMX inhibitor treatment. Competitive advantage and accumulation of b-Catenin upon Mdmx overexpression were still seen upon crossing into a p53-/- background, demonstrating that Mdmx-driven WNT/b-Catenin activation and its functional consequences are, at least part, independent of p53. Lastly, upregulation of WNT/b-Catenin pathways correlated with MDMX overexpression in large cohorts of patients with MDS and was associated with increased risk of progression to AML and poor survival. Our work identifies MDMX overexpression as a pervasive preleukemic-to-AML switch mechanism effective in different genetically-driven disease subtypes, and reveals WNT/b-Catenin as a novel, non-canonical p53-independent MDMX-driven pathway with therapeutic potential for progression prevention and cancer interception.

Inappropriate localization of proteins can interfere with normal cellular function and drive tumor development. To understand how this contributes to the development of acute myeloid leukemia (AML), we compared the nuclear proteome and transcriptome of AML blasts with normal human CD34+ cells. Analysis of the proteome identified networks and processes that significantly affected transcription regulation including misexpression of 11 transcription factors with seven proteins not previously implicated in AML. Transcriptome analysis identified changes in 40 transcription factors but none of these were predictive of changes at the protein level. The highest differentially expressed protein in AML nuclei compared with normal CD34+ nuclei (not previously implicated in AML) was S100A4. In an extended cohort, we found that over-expression of nuclear S100A4 was highly prevalent in AML (83%; 20/24 AML patients). Knock down of S100A4 in AML cell lines strongly impacted their survival whilst normal hemopoietic stem progenitor cells were unaffected. These data are the first analysis of the nuclear proteome in AML and have identified changes in transcription factor expression or regulation of transcription that would not have been seen at the mRNA level. These data also suggest that S100A4 is essential for AML survival and could be a therapeutic target in AML.

Pu.1 is an ETS family transcription factor (TF) that plays critical roles in erythroid progenitors by promoting proliferation and blocking terminal differentiation. However, the mechanisms controlling expression and down-regulation of Pu.1 during early erythropoiesis have not been defined. In this study, we identify the actions of Runx1 and Pu.1 itself at the Pu.1 gene Upstream Regulatory Element (URE) as major regulators of Pu.1 expression in Burst-Forming Unit erythrocytes (BFUe). During early erythropoiesis, Runx1 and Pu.1 levels decline, and chromatin accessibility at the URE is lost. Ectopic expression of Runx1 or Pu.1, both of which bind the URE, prevents Pu.1 down-regulation and blocks terminal erythroid differentiation, resulting in extensive ex vivo proliferation and immortalization of erythroid progenitors. Ectopic expression of Runx1 in BFUe lacking a URE fails to block terminal erythroid differentiation. Thus, Runx1, acting at the URE, and Pu.1 itself directly regulate Pu.1 levels in erythroid cells, and loss of both factors is critical for Pu.1 down-regulation during terminal differentiation. The molecular mechanism of URE inactivation in erythroid cells through loss of TF binding represents a distinct pattern of Pu.1 regulation from those described in other hematopoietic cell types such as T cells which down-regulate Pu.1 through active repression. The importance of down-regulation of Runx1 and Pu.1 in erythropoiesis is further supported by genome-wide analyses showing that their DNA-binding motifs are highly overrepresented in regions that lose chromatin accessibility during early erythroid development.

This paper gives a detailed analysis of the Cannon{Thurston maps associated to a general class of hyperbolic free group extensions. Let F denote a free groups of nite rank at least 3 and consider a convex cocompact subgroup Out(F), i.e. one for which the orbit map from into the free factor complex of F is a quasi-isometric embedding. The subgroup determines an extension E of F, and the main theorem of Dowdall{Taylor (DT1) states that in this situation E is hyperbolic if and only if is purely atoroidal. Here, we give an explicit geometric description of the Cannon{Thurston maps @F! @E for these hyperbolic free group extensions, the existence of which follows from a general result of Mitra. In particular, we obtain a uniform bound on the multiplicity of the Cannon{Thurston map, showing that this map has multiplicity at most 2 rank(F). This theorem generalizes the main result of Kapovich and Lustig (KL5) which treats the special case where is innite cyclic. We also answer a question of Mahan Mitra by producing an explicit example of a hyperbolic free group extension for which the natural map from the boundary of to the space of laminations of the free group (with the Chabauty topology) is not continuous.

Male Red-winged Blackbirds (Agelaius phoeniceus) have red epaulets that are used as badges for intrasexual competition for territory acquisition and defense. Red epaulets reflect very litt...

We prove an effective version of a theorem relating curve complex distance to electric distance in hyperbolic 3-manifolds, up to errors that are polynomial in the complexity of the underlying surface. We use this to give an effective proof of a result regarding maps between curve complexes of surfaces induced by finite covers. As applications, we effectively relate the electric circumference of a fibered manifold to the curve complex translation length of its monodromy, and we give quantitative bounds on virtual specialness for cube complexes dual to curves on surfaces., Several edits including a streamlined proof of Lemma 5.2. Accepted for publication by the Journal of Topology

Every pseudo-Anosov mapping class $\varphi$ defines an associated veering triangulation $\tau_\varphi$ of a punctured mapping torus. We show that generically, $\tau_\varphi$ is not geometric. Here, the word "generic" can be taken either with respect to random walks in mapping class groups or with respect to counting geodesics in moduli space. Tools in the proof include Teichm\"uller theory, the Ending Lamination Theorem, study of the Thurston norm, and rigorous computation., Comment: 38 pages, 9 figures. To appear in Groups, Geometry, and Dynamics

Patient-derived xenograft (PDX) and CTC-derived explant (CDX) models are powerful methods for the study of human disease. In cancer research, these methods have been applied to multiple questions including the study of metastatic progression, genetic evolution and therapeutic drug responses. Since PDX and CDX models can recapitulate the highly heterogeneous characteristics of a patient tumor, as well as their response to chemotherapy, there is considerable interest in combining them with next-generation sequencing (NGS) in order to monitor the genomic, transcriptional, and epigenetic changes that accompany oncogenesis. When used for this purpose, their reliability is highly dependent on being able to accurately distinguish between sequencing reads that originate from the host, and those that arise from the xenograft itself. Here we demonstrate that failure to correctly identify contaminating host reads, when analyzing DNA- and RNA-sequencing (DNA-Seq and RNA-Seq) data from PDX and CDX models is a major confounding factor that can lead to incorrect mutation calls and a failure to identify canonical mutation signatures associated with tumorigenicity. In addition, a highly sensitive algorithm and open source software tool for identifying and removing contaminating host sequences is described. Importantly, when applied to PDX and CDX models of melanoma, these data demonstrate its utility as a sensitive and selective tool for the correction of PDX- and CDX-derived whole exome and RNA-Seq data. Fil: Khandelwal, Garima. University of Manchester; Reino Unido Fil: Girotti, Maria Romina. University of Manchester; Reino Unido. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Smowton, Christopher. University of Manchester; Reino Unido Fil: Taylor, Sam. University of Manchester; Reino Unido Fil: Wirth, Christopher. University of Manchester; Reino Unido Fil: Dynowski, Marek. University of Manchester; Reino Unido Fil: Frese, Kristopher K.. University of Manchester; Reino Unido Fil: Brady, Ged. University of Manchester; Reino Unido Fil: Dive, Caroline. University of Manchester; Reino Unido Fil: Marais, Richard. University of Manchester; Reino Unido Fil: Miller, Crispin. University of Manchester; Reino Unido

We describe an approach to inhibit chemotherapy-induced myelosuppression. We found that short-term exposure of mice to the FLT3 inhibitor quizartinib induced the transient quiescence of multipotent progenitors (MPPs). This property of quizartinib conferred marked protection to MPPs in mice receiving fluorouracil or gemcitabine. The protection resulted in the rapid recovery of bone marrow and blood cellularity, thus preventing otherwise lethal myelosuppression. A treatment strategy involving quizartinib priming that protected wild-type bone marrow progenitors, but not leukemic cells, from fluorouracil provided a more effective treatment than conventional induction therapy in mouse models of acute myeloid leukemia. This strategy has the potential to be extended for use in other cancers where FLT3 inhibition does not adversely affect the effectiveness of chemotherapy. Thus, the addition of quizartinib to cancer treatment regimens could markedly improve cancer patient survival and quality of life.

In this note, we generalize a theorem of Juan Souto on rank and Nielsen equivalence in the fundamental group of a hyperbolic fibered 3-manifold to a large class of hyperbolic group extensions. This includes all hyperbolic extensions of surfaces groups as well as hyperbolic extensions of free groups by convex cocompact subgroups of Out$(F_n)$., Comment: v2: 10 pages. Minor updates to incorporate referee comments. Added counter-example demonstrating necessity of torsion free hypothesis. Final version; accepted for publication in the International Journal of Algebra and Computation (IJAC)

FLT3 internal tandem duplication (FLT3ITD) are common mutations in acute myeloid leukemia (AML) associated with poor patient prognosis. Although new generation FLT3 tyrosine kinase inhibitors (TKI) have shown promising results, the outcome of FLT3ITD AML patients remains poor and demands the identification of novel, specific and validated therapeutic targets for this highly aggressive AML subtype. Utilizing an unbiased genome-wide CRISPR/Cas9 screen, we identify GLS, the first enzyme in glutamine metabolism, as synthetically lethal with FLT3-TKI treatment. Using complementary metabolomic and gene-expression analysis, we demonstrate that glutamine metabolism, through its ability to support both mitochondrial function and cellular redox metabolism, becomes a metabolic dependency of FLT3ITD AML, specifically unmasked by FLT3-TKI treatment. We extend these findings to AML subtypes driven by other tyrosine kinase (TK) activating mutations, and validate the role of GLS as a clinically actionable therapeutic target in both primary AML and in vivo models. Our work highlights the role of metabolic adaptations as a resistance mechanism to several TKI, and suggests glutaminolysis as a therapeutically targetable vulnerability when combined with specific TKI in FLT3ITD and other TK activating mutation driven leukemias.

We prove a central limit theorem for the length of closed geodesics in any compact orientable hyperbolic surface. In the special case of a hyperbolic pair of pants, this settles a conjecture of Chas–Li–Maskit.

We prove that stability -- a strong quasiconvexity property -- pulls back under proper actions on proper metric spaces. This result has several applications, including that convex cocompact subgroups of both mapping class groups and outer automorphism groups of free groups are stable. We also characterize stability in relatively hyperbolic groups whose parabolic subgroups have linear divergence., 21 pages, 1 figure. Update includes some corrections and revisions. To appear in JLMS

We show that a finite collection of stable subgroups of a finitely generated group has finite height, finite width and bounded packing. We then use knowledge about intersections of conjugates to characterize finite families of quasimorphisms on hyperbolically embedded subgroups that can be to simultaneously extended to the ambient group., Comment: Version 2 has a new title, added applications to bounded cohomology, and two additional authors. 16 pages

We show that the largest subsurface projection distance between a marking and its image under the nth step of a random walk grows logarithmically in n, with probability approaching 1 as n tends to infinity. Our setup is general and also applies to (relatively) hyperbolic groups and to $\mathrm{Out}(F_n)$. We then use this result to prove Rivin's conjecture that for a random walk $(w_n)$ on the mapping class group, the shortest geodesic in the hyperbolic mapping torus $M_{w_n}$ has length on the order of $1/ \log^2(n)$., Comment: Version 2: Added application to systole of random mapping tori. 19 pages. To appear in Math. Res. Lett

Let $G \curvearrowright X$ be a nonelementary action by isometries of a hyperbolic group $G$ on a hyperbolic metric space $X$. We show that the set of elements of $G$ which act as loxodromic isometries of $X$ is generic. That is, for any finite generating set of $G$, the proportion of $X$--loxodromics in the ball of radius $n$ about the identity in $G$ approaches $1$ as $n \to \infty$. We also establish several results about the behavior in $X$ of the images of typical geodesic rays in $G$; for example, we prove that they make linear progress in $X$ and converge to the Gromov boundary $\partial X$. Our techniques make use of the automatic structure of $G$, Patterson--Sullivan measure on $\partial G$, and the ergodic theory of random walks for groups acting on hyperbolic spaces. We discuss various applications, in particular to Mod(S), Out($F_N$), and right--angled Artin groups.

We show for an alternating knot the minimal boundary slope of an essential spanning surface is given by the signature plus twice the minimum degree of the Jones polynomial and the maximal boundary slope of an essential spanning surface is given by the signature plus twice the maximum degree of the Jones polynomial. For alternating Montesinos knots, these are the minimal and maximal boundary slopes.

A method is described for synthesizing asynchronous circuits based on the Handshake Circuit paradigm but employing a data-driven, rather than a control-driven, style. This approach attempts to combine the performance advantages of data-driven asynchronous design styles with the handshake circuit style of construction used in existing syntax-directed synthesis. The method is demonstrated on a significant design-a 32-bit microprocessor. This example shows that the data-driven circuit style provides better performance than control-driven synthesized circuits. This paper extends previous reported work by illustrating how conditional execution, oft-cited as a problem for data-driven descriptions, is handled within the system, and by a more detailed analysis of the design example.

Dasatinib is an orally available broad-spectrum tyrosine kinase inhibitor that is widely used to treat chronic myeloid leukemia. It is also in clinical trials for the treatment of other malignancies, including solid tumors. Despite its wide use, little is known of its effects on normal hematopoietic stem and progenitor cells. Here, we study wild-type mice dosed with dasatinib and find that it causes the transient induction of proliferation of quiescent hematopoietic stem cells (HSCs). This finding was unexpected given the ability of dasatinib to inhibit c-Kit signaling and promote cell cycle arrest in many cell types. The transient induction of HSC proliferation in dasatinib-dosed mice coincided with a marked induction in the expression of Sca-1 and phospho-S6. Also evident at this time was a rapid but transient loss of lineage-committed hematopoietic progenitors that express high levels of c-Kit and the induction of stem cell factor in the serum. These findings suggest that activation of quiescent HSCs is part of a rapid rescue response that restores hematopoietic progenitors to pretreatment levels. This restoration coincides with HSCs returning to quiescence, and the expression of Sca-1 and phospho-S6 reverting to pre-treatment levels, even though dasatinib dosing is maintained. These data suggest that equilibrium is reached between the opposing forces of dasatinib and hematopoietic growth factors. The transient induction of HSC proliferation provided a window of opportunity whereby these cells became sensitive to killing by the cytotoxic drug 5-fluorouracil.

We show that strongly contracting geodesics in Outer space project to parameterized quasigeodesics in the free factor complex. This result provides a converse to a theorem of Bestvina--Feighn, and is used to give conditions for when a subgroup of $\mathrm{Out}(\mathbb{F})$ has a quasi-isometric orbit map into the free factor complex. It also allows one to construct many new examples of strongly contracting geodesics in Outer space., v3: 20 pages. minor corrections and changes based on referee comments. Final version; accepted for publication in MathZ

In this note, we prove that a random extension of either the free group $F_N$ of rank $N\ge3$ or of the fundamental group of a closed, orientable surface $S_g$ of genus $g\ge2$ is a hyperbolic group. Here, a random extension is one corresponding to a subgroup of either Out$(F_N)$ or Mod$(S_g)$ generated by $k$ independent random walks. Our main theorem has several applications, including that a random subgroup of a weakly hyperbolic group is free and undistorted., Comment: 13 pages

We introduce a strong notion of quasiconvexity in finitely generated groups, which we call stability. Stability agrees with quasiconvexity in hyperbolic groups and is preserved under quasi-isometry for finitely generated groups. We show that the stable subgroups of mapping class groups are precisely the convex cocompact subgroups. This generalizes a well-known result of Behrstock and is related to questions asked by Farb-Mosher and Farb., Comment: 15 pages, 1 figure

Protecting an endangered and highly poached species can conflict with providing an open and ecologically connected landscape for coexisting species. In Kenya, about half of the black rhino (Diceros bicornis) live in electrically fenced private conservancies. Purpose-built fence-gaps permit some landscape connectivity for elephant while restricting rhino from escaping. We monitored the usage patterns at these gaps by motion-triggered cameras and found high traffic volumes and predictable patterns of prey movement. The prey-trap hypothesis (PTH) proposes that predators exploit this predictable prey movement. We tested the PTH at two semi-porous reserves using two different methods: a spatial analysis and a temporal analysis. Using spatial analysis, we mapped the location of predation events with GPS and looked for concentration of kill sites near the gaps as well as conducting clustering and hot spot analysis to determine areas of statistically significant predation clustering. Using temporal analysis, we examined the time lapse between the passage of prey and predator and searched for evidence of active prey seeking and/or predator avoidance. We found no support for the PTH and conclude that the design of the fence-gaps is well suited to promoting connectivity in these types of conservancies.

Functionally critical decreases in levels or activity of the ETS family transcription factor PU.1 are present in approximately 2/3 of patients with acute myeloid leukemia (AML), across different AML subtypes (Sive, Leukemia 2016) including at the stem cell level (Steidl, Nat Genet 2006; Will, Nat Med 2015). Thus, targeting PU.1 could be an appealing option for treatment. As complete loss of PU.1 leads to stem cell failure (Iwasaki, Blood 2005), we hypothesized that PU.1 inhibition could eradicate leukemic cells harboring already low levels of PU.1, with modest effects on normal cells. We initially tested this hypothesis using 3 different shRNAs, and found that PU.1 inhibition led to a significant decrease in proliferation and clonogenicity, and increased apoptosis of mouse and human leukemic cell lines with low PU.1 levels, as well as the majority of primary human AML cells tested. We demonstrated that these effects were indeed due to decreased PU.1 levels by retroviral add-back experiments. The direct pharmacologic targeting of transcription factors has proven challenging in the past. Besides the core ETS binding motif (GGAA) in the DNA major groove, PU.1 binding to chromatin depends on additional minor groove contacts enriched for AT nucleotides upstream of the ETS motif, which determine selectivity for PU.1. Using an integrated screening strategy utilizing biosensor surface plasmon resonance, DNA footprinting, and cell-based dual-color PU.1 reporter assays, we developed novel small molecules of the heterocyclic diamidine family acting as first-in-class PU.1 inhibitors. Targeted occupancy by our compounds in the minor groove induces perturbations in DNA conformation that are transmitted to the PU.1 site in the major groove and thus inhibits PU.1 binding via an allosteric mechanism. Consistent with this, the inhibitory effects were selective for PU.1 versus other ETS transcription factors. Treatment with 3 different compounds led to cell growth inhibitory effect with respect to PU.1 level and preferentially affects PU.1low AML cells. Similarly to what we observed with shRNAs, treatment with our novel inhibitors led to decreased proliferation and colony forming capacity, increased apoptosis, and disrupted serial replating capacity of PU.1low AML cells and a majority of primary AML cell samples. Targeted ChIP and expression analysis showed that the compounds disrupt PU.1-promoter interaction and lead to downregulation of canonical PU.1 transcriptional targets in AML cells, confirming on-target activity in AML cells. Genome-wide analysis showed highly significant enrichment of known transcriptional targets of PU.1, and selectivity over genes regulated by other ETS family members. Comparison with published transcriptomic and PU.1 ChIP-seq data sets, as well as ARACNe analysis of the PU.1 regulon in primary AML cells, demonstrated that the inhibitors antagonize PU.1-regulated pathways at a genome-wide level. ChIP-seq performed in PU.1low AML cells confirmed a genome-wide decrease of PU.1 peaks after treatment and provides novel insight into the molecular mechanisms mediating the anti-leukemic effects of pharmacological PU.1 inhibition. To test the effects of PU.1 inhibition on normal hematopoiesis, we treated normal hematopoietic stem/progenitors cells (HSPC) in colony forming assays and saw decreased production of mature granulo-monocytic cells, consistent with PU.1's known role in this lineage. However, this effect was reversible upon drug removal, and serial replating capacity was not affected suggesting no significant effects on more immature HSPC. Congenic transplantation assays of treated normal bone marrow cells led to no change in myeloid and T-cells and only a modest decrease in B-cell numbers. Lastly, in vivo treatment with PU.1 inhibitors in mouse and human AML (xeno)transplantation models significantly decreased tumor burden and increased survival. To conclude, our study provides proof-of-principle for PU.1 inhibition as a novel therapeutic strategy in AML. Furthermore, we present the development of first-in-class PU.1 inhibitors acting via an allosteric minor groove-mediated mechanism. Our work shows that the specific pharmacological targeting of the DNA interaction of transcription factors such as PU.1 is feasible in principle, and may open the way for targeting of other transcription factors through minor groove-directed approaches. Disclosures Will: Novartis Pharmaceuticals: Consultancy, Research Funding. Steidl: Celgene: Consultancy; Aileron Therapeutics: Consultancy, Research Funding; Novartis: Research Funding; GlaxoSmithKline: Research Funding; Bayer Healthcare: Consultancy.

Dasatinib inhibits B-cell receptor–Abelson murine leukemia viral oncogene homologue 1, Src, and other tyrosine kinases. Few studies have addressed the impact of dasatinib on normal blood cells, especially in vivo. Here we show that dasatinib leads to a reduced number of human CD19 + peripheral B cells owing to a strong induction of apoptosis. In contrast, no similar effect on T-cell viability was observed. However, dasatinib induced a comparable broad inhibition of the early events of B- and T-cell receptor signaling. Furthermore, dasatinib was shown to be a more pronounced inhibitor of both basal and B-cell receptor–induced activity of Bruton's tyrosine kinase and PLCγ2 compared with the more specific Bruton's tyrosine kinase inhibitor ibrutinib. Human progenitor B cells from the pre-B stage were sensitive to dasatinib. In an in vivo murine model, dasatinib reduced B-lineage cells in the bone marrow with a marked effect on the pre-B subpopulation. Dasatinib led to a reduced spleen size, with a loss of large immature transitional immunoglobulin M + /immunoglobulin D − B cells and a reduction in germinal center B cells. Dasatinib caused a marked loss of thymocytes without affecting myeloid lineage cells or hematopoietic progenitors. This study reveals important side effects of dasatinib with specific loss of activated B and thymocyte populations, which may have an impact during long-term treatment.

Given a finitely generated subgroup $\Gamma \le \mathrm{Out}(\mathbb{F})$ of the outer automorphism group of the rank $r$ free group $\mathbb{F} = F_r$, there is a corresponding free group extension $1 \to \mathbb{F} \to E_{\Gamma} \to \Gamma \to 1$. We give sufficient conditions for when the extension $E_{\Gamma}$ is hyperbolic. In particular, we show that if all infinite order elements of $\Gamma$ are atoroidal and the action of $\Gamma$ on the free factor complex of $\mathbb{F}$ has a quasi-isometric orbit map, then $E_{\Gamma}$ is hyperbolic. As an application, we produce examples of hyperbolic $\mathbb{F}$-extensions $E_{\Gamma}$ for which $\Gamma$ has torsion and is not virtually cyclic. The proof of our main theorem involves a detailed study of quasigeodesics in Outer space that make progress in the free factor complex. This may be of independent interest., Comment: 50 pages. Minor changes and other updates to incorporate referee comments. Final version; accepted for publication in Geometry & Topology

We extend some results of [BF12] on subfactor projections to show that the projection of a free factor B to the free factor complex of the free factor A is well-defined with uniformly bound diameter, unless either A is contained in B or A and B are vertex stabilizers of a single splitting of F_n, i.e. they are disjoint. These projections are shown to satisfy properties analogous to subsurface projections, and we give as an application a construction of fully irreducible outer automorphisms using the Bounded Geodesic Image Theorem., 13 pages

Let $S_{g,p}$ denote the genus $g$ orientable surface with $p \ge 0$ punctures, and let $\omega(g,p)= 3g+p-4$. We prove the existence of infinitely long geodesic rays $\left\{v_{0},v_{1}, v_{2}, ...\right\}$ in the curve graph satisfying the following optimal intersection property: for any natural number $k$, the endpoints $v_{i},v_{i+k}$ of any length $k$ subsegment intersect $O(\omega^{k-2})$ times. By combining this with work of the first author, we answer a question of Dan Margalit., Comment: 13 pages, 6 figures

We show that for a surface S, the subgraph of the pants graph determined by fixing a collection of curves that cut S into pairs of pants, once-punctured tori, and four-times-punctured spheres is totally geodesic. The main theorem resolves a special case of a conjecture made by Aramayona, Parlier, and Shackleton and has the implication that an embedded product of Farey graphs in any pants graph is totally geodesic. In addition, we show that a pants graph contains a convex n-flat if and only if it contains an n-quasi-flat., v2: 25 pages, 16 figures. Completely rewritten, several figures added for clarity