Ikaros fingers on lymphocyte differentiation

The Ikaros family of DNA binding proteins are critical regulators of lymphocyte differentiation. In multipotent hematopoietic progenitors, Ikaros supports transcriptional priming of genes promoting lymphocyte differentiation. Ikaros targets the Nucleosome Remodeling Deacetylase complex (NuRD) to lymphoid lineage genes, thereby increasing chromatin accessibility and transcriptional priming. After lymphoid lineage specification, Ikaros expression is raised to levels characteristic of intermediate B cell and T cell precursors, which is necessary to support maturation and prevent leukemogenesis. Loss of Ikaros in T cell precursors allows the NuRD complex to repress lymphocyte genes and extends its targeting to genes that support growth and proliferation, causing their activation and triggering a cascade of events that leads to leukemogenesis. Loss of Ikaros in B cell precursors blocks differentiation and perpetuates stromal adhesion by enhancing integrin signaling. The combination of integrin and cytokine signaling in Ikaros-deficient pre-B cells promotes their survival and self-renewal. The stages of lymphocyte differentiation that are highly dependent on Ikaros are underscored by changes in Ikaros transcription, supported by a complex network of stage-specific regulatory networks that converge upon the Ikzf1 locus. It is increasingly apparent that understanding the regulatory networks that operate upstream and downstream of Ikaros is critical not only for our understanding of normal lymphopoiesis, but also in placing the right finger on the mechanisms that support hematopoietic malignancies in mouse and human

An Ikaros Promoter Element with Dual Epigenetic and Transcriptional Activities

Ikaros DNA binding factor plays critical roles in lymphocyte development. Changes in Ikaros expression levels during lymphopoiesis are controlled by redundant but also unique regulatory elements of its locus that are critical for this developmental process. We have recently shown that Ikaros binds its own locus in thymocytes in vivo. Here, we evaluated the role of an Ikaros binding site within its major lympho-myeloid promoter. We identified an Ikaros/Ets binding site within a promoter sub-region that was highly conserved in mouse and human. Deletion of this binding site increased the percentage of the reporter-expressing mouse lines, indicating that its loss provided a more permissive chromatin environment. However, once transcription was established, the lack of this site decreased transcriptional activity. These findings implicate a dual role for Ikaros/Ets1 binding on Ikzf1 expression that is exerted at least through its promoter

New Stepwedge for Bootstrap Sensitometry in Medical Radiography

A new stepwedge has been developed for bootstrap sensitometry, which is used for determining the characteristic curve of radiographic screen/film systems. Since each step of the stepwedge is separated by the lead septum, flare generated under one step will not penetrate the region of another step. The metal of the stepwedge can be changed with various materials. Characteristic curves obtained with new stepwedge bootstrap (NSB) and conventional stepwedge bootstrap (CSB) techniques were compared with those obtained for screen/film systems with inverse sqare sensitometry. The inverse square method was used as the reference standard for accuracy. The NSB method provided characteristic curves that agreed well with those obtained using the inverse square method. The slope of the curve in the CSB method was lower than that in the inverse square or NBS method, because of scatter radiation. The shape of the characteristic curve from the NSB method did not vary with the materials of step metals (aluminum and copper). The cumulative error involved when using the bootstrap method is also discussed

Influence on the character of films used for density control of an automatic processor by the difference in the way of preserving

自動現像機の濃度管理に用いられるフィルムの保存については，種々の報告がなされており，最良の方法が確立されているとは言い難い。さらに，レギュラフィルムとオルソフィルムの保存方法の違いによる濃度管理への影響，あるいはこれらの比較については，これまでほとんど報告されていない。本論文では，われわれは，レギュラフィルム，フジnew-RXとオルソフィルム，コニカSR-G，SR－VおよびSR-Hを用い，開封後，室温，冷蔵，冷凍の3種類の保存方法を採用した。われわれは，35週間3種類の保存方法で4種類 のフィルムのgross　fog，speed indexとaverage gradientへの影響を調査した。その結果レギュラフィルムは，3種類の保存方法による差が認められなかった。オルソフィルムでは，冷蔵・冷凍保存に大きな特性変動が認められた。その原因として，オルソフィルムは増感色素が結露に対して敏感に影響を受けたと考えられた。Though there are various reports on the way of preserving films used for density control of the automatic processor, it is hard to say that the best method isestablished. In　addition, the influences on density control or their comparisons by the difference way of preserving film between regular and orthochromatic films have been hardly reported until now. In this paper, three ways of preserving films were employed, which were at a room temperature, in a refrigeration and in a freezer after the films were opened, using a regular film, Fuji new-RX and three orthochromatic films, Konica SR-G, SR-V and SR-H. We investigated the influences on gross fog, speed index and average gradient of four films on three different ways of preserving film for 35 weeks. As a result, new-RX film wasn't influenced by the ways of the preservation. On the other hand, the orthochromatic films were so influences sensitively that the films couldn't be used for density control. It was considered that sensitizing dye stuff got denatured by dew condensation

Promoter Decommissioning by the NuRD Chromatin Remodeling Complex Triggers Synaptic Connectivity in the Mammalian Brain

SummaryPrecise control of gene expression plays fundamental roles in brain development, but the roles of chromatin regulators in neuronal connectivity have remained poorly understood. We report that depletion of the NuRD complex by in vivo RNAi and conditional knockout of the core NuRD subunit Chd4 profoundly impairs the establishment of granule neuron parallel fiber/Purkinje cell synapses in the rodent cerebellar cortex in vivo. By interfacing genome-wide sequencing of transcripts and ChIP-seq analyses, we uncover a network of repressed genes and distinct histone modifications at target gene promoters that are developmentally regulated by the NuRD complex in the cerebellum in vivo. Finally, in a targeted in vivo RNAi screen of NuRD target genes, we identify a program of NuRD-repressed genes that operate as critical regulators of presynaptic differentiation in the cerebellar cortex. Our findings define NuRD-dependent promoter decommissioning as a developmentally regulated programming mechanism that drives synaptic connectivity in the mammalian brain

Harnessing of the Nucleosome Remodeling Deacetylase complex controls lymphocyte development and prevents leukemogenesis

Cell fate decisions depend on the interplay between chromatin regulators and transcription factors. Here we show that activity of the Mi-2β nucleosome remodeling and deacetylase (NuRD) complex was controlled by the Ikaros family of lymphoid-lineage determining proteins. Ikaros, an integral component of the NuRD complex in lymphocytes, tethered this complex to active lymphoid differentiation genes. Loss in Ikaros DNA binding activity caused a local increase in Mi-2β chromatin remodeling and histone deacetylation and suppression of lymphoid gene expression. The NuRD complex also redistributed to transcriptionally poised non-Ikaros gene targets, involved in proliferation and metabolism, inducing their reactivation. Thus, release of NuRD from Ikaros regulation blocks lymphocyte maturation and mediates progression to a leukemic state by engaging functionally opposing epigenetic and genetic networks

Physical Image Quality of Rapid Processing Films

The imaging properties of Konica "new" SR series films and "old" MG-SR series films for rapid processing were measured in combination with green-emitting intensifying screens. The measured image quality prarameters include: characteristic curve, modulation transfer function (MTF) and Wiener spectrum. The relative speeds of the new SR series systems were slower than those of the old MG-SR series systems from 10 to 25%. The MTF's of the new systems were generally comparable to those of the old systems, except that SR-G system was improved with an increase of approximately 10%. In the Wiener spectrum measurement, however, the new systems showed from 10 to 35% decrease in low-frequency noise

Awakening lineage potential by Ikaros-mediated transcriptional priming

Bioinformatic studies on a revised hierarchy of hematopoietic progenitors have provided a genome-wide view of lineage-affiliated transcriptional programs directing early hematopoiesis. Unexpectedly, lymphoid, myeloid, and erythroid gene expression programs were primed with similar frequency at the multipotent progenitor stage indicating a stochastic nature to this process. Multilineage transcriptional priming is quickly resolved upon erythroid lineage restriction with both lymphoid and myeloid transcriptional programs rapidly extinguished. However, expression of lymphoid and myeloid factors remains active past nominal lymphoid and myeloid lineage restrictions, revealing a common genetic network utilized by both pathways. Priming and resolution of multilineage potential is dependent on the activity of the DNA binding factor Ikaros. Ikaros primes the lymphoid transcriptional program in the HSC and represses the stem cell and other disparate transcriptional programs downstream of the HSC. Loss of Ikaros removes the lymphoid leg of the immune system and may confer aberrant self-renewing properties to myeloid progenitors