Nfix expression critically modulates early B lymphopoiesis and myelopoiesis

The commitment of stem and progenitor cells toward specific hematopoietic lineages is tightly controlled by a number of transcription factors that regulate differentiation programs via the expression of lineage restricting genes. Nuclear factor one (NFI) transcription factors are important in regulating hematopoiesis and here we report an important physiological role of NFIX in B- and myeloid lineage commitment and differentiation. We demonstrate that NFIX acts as a regulator of lineage specification in the haematopoietic system and the expression of Nfix was transcriptionally downregulated as B cells commit and differentiate, whilst maintained in myeloid progenitor cells. Ectopic Nfix expression in vivo blocked early B cell development stage, coincident with the stage of its downregulation. Furthermore, loss of Nfix resulted in the perturbation of myeloid and lymphoid cell differentiation, and a skewing of gene expression involved in lineage fate determination. Nfix was able to promote myeloid differentiation of total bone marrow cells under B cell specific culture conditions but not when expressed in the hematopoietic stem cell (HSPC), consistent with its role in HSPC survival. The lineage choice determined by Nfix correlated with transcriptional changes in a number of genes, such as E2A, C/EBP, and Id genes. These data highlight a novel and critical role for NFIX transcription factor in hematopoiesis and in lineage specification

Effect of Mill Type on Morphology of AA6013 Aluminium Powder

ABSTRACTIn conventional recycling method, metal chips are cast after pressing and melting in electric arc furnace. Material loss occurs during the recycling from liquid metal due to the several reasons. Direct recycling method which produces the aluminium powder from aluminium chips using mechanical mill can be an alternative to conventional recycling method. Thus material and energy losses, and labour cost will be reduced by direct recycling method without melting.In this study, the particle morphology of powder direct recycled from AA6013 aluminium alloy chips with cryogenic, disc and ball type grinders is investigated. Mechanical milling resulted flaky and irregular shaped AA6013 particles. It was ascertained that the chips did not break sufficiently in despite of the long duration milling mechanisms by ball mill. Cryogenic mill provides the energy required for milling mechanisms to act. Disc mill has the highest impact energy was determined. Consequently, efficiency of ball mill is lower than the efficiency of cryogenic and disc type mills. Shape factors of powders produced with ball and cryogenic mills were found greater than that of the powder produced by disc mill. Disc mill has the most efficient and effective impact energy which produces the smaller particles per minute, was determined.Keywords: Direct recycling method, powder production, scrap chips, aluminium alloy.

Abnormal development of forebrain midline glia and commissural projections in Nfia knock-out mice

Nuclear factor I (NFI) genes are expressed in multiple organs throughout development (Chaudhry et al., 1997; for review, see Gronostajski, 2000). All four NFI genes are expressed in embryonic mouse brain, with Nfia, Nfib, and Nfix being expressed highly in developing cortex (Chaudhry et al., 1997). Disruption of the Nfia gene causes agenesis of the corpus callosum (ACC), hydrocephalus, and reduced GFAP expression (das Neves et al., 1999). Three midline structures, the glial wedge, glia within the indusium griseum, and the glial sling are involved in development of the corpus callosum (Silver et al., 1982; Silver and Ogawa, 1983; Shu and Richards, 2001). Because Nfia(-/-) mice show glial abnormalities and ACC, we asked whether defects in midline glial structures occur in Nfia(-/-) mice. NFI-A protein is expressed in all three midline populations. In Nfia(-/-) mice sling cells are generated but migrate abnormally into the septum and do not form a sling. Glia within the indusium griseum and the glial wedge are greatly reduced or absent and consequently Slit2 expression is also reduced. Although callosal axons approach the midline, they fail to cross and extend aberrantly into the septum. The hippocampal commissure is absent or reduced, whereas the ipsilaterally projecting perforating axons (Hankin and Silver, 1988; Shu et al., 2001) appear relatively normal. These results support an essential role for midline glia in callosum development and a role for Nfia in the formation of midline glial structures

nfi-1 affects behavior and life-span in C. elegans but is not essential for DNA replication or survival

BACKGROUND: The Nuclear Factor I (one) (NFI) family of transcription/replication factors plays essential roles in mammalian gene expression and development and in adenovirus DNA replication. Because of its role in viral DNA replication NFI has long been suspected to function in host DNA synthesis. Determining the requirement for NFI proteins in mammalian DNA replication is complicated by the presence of 4 NFI genes in mice and humans. Loss of individual NFI genes in mice cause defects in brain, lung and tooth development, but the presence of 4 homologous NFI genes raises the issue of redundant roles for NFI genes in DNA replication. No NFI genes are present in bacteria, fungi or plants. However single NFI genes are present in several simple animals including Drosophila and C. elegans, making it possible to test for a requirement for NFI in multicellular eukaryotic DNA replication and development. Here we assess the functions of the single nfi-1 gene in C. elegans. RESULTS: C. elegans NFI protein (CeNFI) binds specifically to the same NFI-binding site recognized by vertebrate NFIs. nfi-1 encodes alternatively-spliced, maternally-inherited transcripts that are expressed at the single cell stage, during embryogenesis, and in adult muscles, neurons and gut cells. Worms lacking nfi-1 survive but have defects in movement, pharyngeal pumping and egg-laying and have a reduced life-span. Expression of the muscle gene Ce titin is decreased in nfi-1 mutant worms. CONCLUSION: NFI gene function is not needed for survival in C. elegans and thus NFI is likely not essential for DNA replication in multi-cellular eukaryotes. The multiple defects in motility, egg-laying, pharyngeal pumping, and reduced lifespan indicate that NFI is important for these processes. Reduction in Ce titin expression could affect muscle function in multiple tissues. The phenotype of nfi-1 null worms indicates that NFI functions in multiple developmental and behavioral systems in C. elegans, likely regulating genes that function in motility, egg-laying, pharyngeal pumping and lifespan maintenance

Nuclear factor I-A represses expression of the cell adhesion molecule L1

<p>Abstract</p> <p>Background</p> <p>The neural cell adhesion molecule L1 plays a crucial role in development and plasticity of the nervous system. Neural cells thus require precise control of L1 expression.</p> <p>Results</p> <p>We identified a full binding site for nuclear factor I (NFI) transcription factors in the regulatory region of the mouse <it>L1 </it>gene. Electrophoretic mobility shift assay (EMSA) showed binding of nuclear factor I-A (NFI-A) to this site. Moreover, for a brain-specific isoform of NFI-A (NFI-A bs), we confirmed the interaction <it>in vivo </it>using chromatin immunoprecipitation (ChIP). Reporter gene assays showed that in neuroblastoma cells, overexpression of NFI-A bs repressed L1 expression threefold.</p> <p>Conclusion</p> <p>Our findings suggest that NFI-A, in particular its brain-specific isoform, represses <it>L1 </it>gene expression, and might act as a second silencer of L1 in addition to the neural restrictive silencer factor (NRSF).</p

Characterization of the austenite recrystallization by comparing double deformation and stress relaxation tests

A high amount of deformation below the non-recrystallization temperature (T-nr) is a common industrial practice to achieve a good combination of toughness and strength in microalloyed steels. To combine the industrially relevant optimum combination of high productivity and product quality, an accurate knowledge of T-nr and the recrystallization kinetics is required. Although a lot of literature data is available on the recrystallization behaviour of microalloyed steels, correlations are often difficult to be made due to the effect of different experimental setups, types of analysis and test schedules that are used to obtain this data. Although this would significantly improve the knowledge about these steels, so far, no systematic comparison has been presented in literature to correlate the different techniques and methods. In this study, different hot rolling simulation techniques, testing schedules and types of analysis were used to determine the recrystallization kinetics of a microalloyed steel. On the one hand, good agreement was found between the results from different test equipment for the double deformations tests. On the other hand, stress relaxation tests showed accelerated kinetics and appeared to be less effective

Nuclear factor I coordinates multiple phases of cerebellar granule cell development via regulation of cell adhesion molecules

A central question is how various stages of neuronal development are integrated as a differentiation program. Here we show that the nuclear factor I (NFI) family of transcriptional regulators is expressed and functions throughout the postmitotic development of cerebellar granule neurons (CGNs). Expression of an NFI dominant repressor in CGN cultures blocked axon outgrowth and dendrite formation and decreased CGN migration. Inhibition of NFI transactivation also disrupted extension and fasciculation of parallel fibers as well as CGN migration to the internal granule cell layer in cerebellar slices. In postnatal day 17 Nfia-deficient mice, parallel fibers were greatly diminished and disoriented, CGN dendrite formation was dramatically impaired, and migration from the external germinal layer (EGL) was retarded. Axonal marker expression also was disrupted within the EGL of embryonic day 18 Nfib-null mice. NFI regulation of axon extension was observed under conditions of homotypic cell contact, implicating cell surface proteins as downstream mediators of its actions in CGNs. Consistent with this, the cell adhesion molecules ephrin B1 and N-cadherin were identified as NFI gene targets in CGNs using inhibitor and Nfi mutant analysis as well as chromatin immunoprecipitation. Functional inhibition of ephrin B1 or N-cadherin interfered with CGN axon extension and guidance, migration, and dendritogenesis in cell culture as well as in situ. These studies define NFI as a key regulator of postmitotic CGN development, in particular of axon formation, dendritogenesis, and migratory behavior. Furthermore, they reveal how a single transcription factor family can control and integrate multiple aspects of neuronal differentiation through the regulation of cell adhesion molecules

Effect of amorphous silica by rice husk ash on physical properties and microstructures of recycled aluminium chip AA7075

High strength to weight ratio of aluminium reinforced as metal matrix composites is a well known material used in automotive application. The effects of recycled aluminium chips AA7075 with amorphous silica by rice husk ash on the physical properties and microstructure were investigated. Recycled aluminium chip AA7075 was reinforced with agro waste of amorphous silica rice husk ash i. e., 2.5 %, 5 %, 7.5 %, 10 % and 12.5 %. Samples of these metal matrix composites were prepared by cold compaction method due to the lower energies consumption and operating cost compared to conventional recycling by casting. Physical testing of density, apparent porosity, water absorption and hardness tests of the metal matrix composites samples were examined in the current study. The density of metal matrix composites was increased up to 5 % of amorphous silica, and then decreased with increasing mass fraction of amorphous silica. Porosity and water absorption of metal matrix composites were significantly consistent at increasing mass fraction of amorphous silica, while the hardness of metal matrix composites was increased at increasing amorphous silica. Consequently, the microstructures of metal matrix composites were observed via optical microscope to analyze the dispersion of the reinforced composites. The microstructures of metal matrix composites were found non-homogeneous and random distribution of amorphous silica and aluminium chip AA7075 compared to 100 % recycled aluminium chip AA7075. Based on investigation to aluminium reinforced rice husk ash composites, it has good potential to improve the material behavior of metal matrix composites by appropriate composition amorphous silica to composite