Evaluation of argyrophilic nucleolar organizer regions in oral tumor progression

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cooperation between the activin and Wnt pathways in the spatial control of organizer gene expression

The normal expression pattern of the Wnt responsive homeobox gene Siamois is restricted to the dorso-vegetal region of the Xenopus embryo. Because the Wnt signaling pathway (via β-catenin) is active on the entire dorsal side of the early embryo, we have asked why Siamois expression is not seen in the dorsal ectoderm. Only Wnt signaling, via activation of β-catenin, can induce directly Siamois, and signaling via the SMAD1 (BMP2/4) or SMAD2 (activin/Vg-1) pathways cannot. We now directly show that the SMAD2 pathway can cooperate with the Wnt pathway to induce expression of Siamois much more strongly than the Wnt pathway alone, in normal embryos. We demonstrate the significance of this cooperation in normal embryos by blocking the SMAD2 signaling pathway with a dominant negative activin receptor. The activin dominant negative receptor blocks this cooperative effect and reduces the expression of Siamois by threefold in early embryos. Furthermore, we find that this cooperative relationship between the SMAD2 and Wnt pathways is reciprocal. Thus, in normal embryos, the Wnt pathway can enhance induction, by the SMAD 2 pathway, of the organizer genes Gsc and Chd but not the pan-mesodermal marker genes Xbra and Eomes. We conclude that the Wnt and SMAD2 signaling pathways cooperate to induce the expression of Spemann-organizer specific genes and so help to localize their spatial expression.

Siamois is required for formation of Spemann’s organizer

Spemann’s organizer develops in response to dorsal determinants that act via maternal components of the wnt pathway. The function of siamois, a wnt-inducible homeobox gene, in Spemann’s organizer development was examined by fusion of defined transcriptional regulatory domains to the siamois homeodomain. Similar to native siamois, a VP16 activator fusion induced axis formation, indicating that siamois functions as a transcriptional activator in axis induction. Fusion of the engrailed repressor generated a dominant inhibitor that blocked axis induction by Xwnt8, β-catenin, and siamois, and repressed wnt activation of the goosecoid promoter. Dorsal injection of the engrailed-siamois fusion resulted in complete inhibition of dorsal development and organizer gene expression, an effect rescued by siamois, but not by Xwnt8 or β-catenin. Thus, as a zygotic mediator of maternal dorsal signals, siamois function is required for development of Spemann’s organizer.

The Three-dimensional Study of Chromosomes and Upstream Binding Factor-immunolabeled Nucleolar Organizer Regions Demonstrates Their Nonrandom Spatial Arrangement during Mitosis

The volumic rearrangement of both chromosomes and immunolabeled upstream binding factor in entire well-preserved mitotic cells was studied by confocal microscopy. By using high-quality three-dimensional visualization and tomography, it was possible to investigate interactively the volumic organization of chromosome sets and to focus on their internal characteristics. More particularly, this study demonstrates the nonrandom positioning of metaphase chromosomes bearing nucleolar organizer regions as revealed by their positive upstream binding factor immunolabeling. During the complex morphogenesis of the progeny nuclei from anaphase to late telophase, the equal partitioning of the nucleolar organizer regions is demonstrated by quantification, and their typical nonrandom central positioning within the chromosome sets is revealed.

Electron Tomography of Metaphase Nucleolar Organizer Regions: Evidence for a Twisted-Loop Organization

Metaphase nucleolar organizer regions (NORs), one of four types of chromosome bands, are located on human acrocentric chromosomes. They contain r-chromatin, i.e., ribosomal genes complexed with proteins such as upstream binding factor and RNA polymerase I, which are argyrophilic NOR proteins. Immunocytochemical and cytochemical labelings of these proteins were used to reveal r-chromatin in situ and to investigate its spatial organization within NORs by confocal microscopy and by electron tomography. For each labeling, confocal microscopy revealed small and large double-spotted NORs and crescent-shaped NORs. Their internal three-dimensional (3D) organization was studied by using electron tomography on specifically silver-stained NORs. The 3D reconstructions allow us to conclude that the argyrophilic NOR proteins are grouped as a fiber of 60–80 nm in diameter that constitutes either one part of a turn or two or three turns of a helix within small and large double-spotted NORs, respectively. Within crescent-shaped NORs, virtual slices reveal that the fiber constitutes several longitudinally twisted loops, grouped as two helical 250- to 300-nm coils, each centered on a nonargyrophilic axis of condensed chromatin. We propose a model of the 3D organization of r-chromatin within elongated NORs, in which loops are twisted and bent to constitute one basic chromatid coil.

Role of the iroquois3 homeobox gene in organizer formation

In zebrafish, the organizer is thought to consist of two regions, the yolk syncytial layer (YSL) and the shield. The dorsal YSL appears to send signals that affect formation of the shield in the overlying mesendoderm. We show here that a domain of dorsal deep cells located between the YSL and the shield is marked by expression of the iro3 gene. As gastrulation proceeds, the iro3 positive domain involutes and migrates to the animal pole. Iro3 expression is regulated by Nodal and bone morphogenic protein antagonists. Overexpression of iro3 induced ectopic expression of shield-specific genes. This effect was mimicked by an Iro3-Engrailed transcriptional repressor domain fusion, whereas an Iro3-VP16 activator domain fusion behaved as a dominant negative or antimorphic form. These results suggest that Iro3 acts as a transcriptional repressor and further implicate the iro3 gene in regulating organizer formation. We propose that the iro3-expressing dorsal deep cells represent a distinct organizer domain that receives signals from the YSL and in turn sends signals to the forming shield, thereby influencing its expansion and differentiation.

Assignment of Rfp-Y to the chicken major histocompatibility complex/NOR microchromosome and evidence for high-frequency recombination associated with the nucleolar organizer region.

Rfp-Y is a second region in the genome of the chicken containing major histocompatibility complex (MHC) class I and II genes. Haplotypes of Rfp-Y assort independently from haplotypes of the B system, a region known to function as a MHC and to be located on chromosome 16 (a microchromosome) with the single nucleolar organizer region (NOR) in the chicken genome. Linkage mapping with reference populations failed to reveal the location of Rfp-Y, leaving Rfp-Y unlinked in a map containing >400 markers. A possible location of Rfp-Y became apparent in studies of chickens trisomic for chromosome 16 when it was noted that the intensity of restriction fragments associated with Rfp-Y increased with increasing copy number of chromosome 16. Further evidence that Rfp-Y might be located on chromosome 16 was obtained when individuals trisomic for chromosome 16 were found to transmit three Rfp-Y haplotypes. Finally, mapping of cosmid cluster III of the molecular map of chicken MHC genes (containing a MHC class II gene and two rRNA genes) to Rfp-Y validated the assignment of Rfp-Y to the MHC/NOR microchromosome. A genetic map can now be drawn for a portion of chicken chromosome 16 with Rfp-Y, encompassing two MHC class I and three MHC class II genes, separated from the B system by a region containing the NOR and exhibiting highly frequent recombination.

The Army of the American Revolution and its organizer ...

Life and work of Gen. von Steuben.

The use of a computer graphic organizer for persuasive composition writing by Hispanic students with specific learning disabilities

Many culturally and linguistically diverse (CLD) students with specific learning disabilities (SLD) struggle with the writing process. Particularly, they have difficulties developing and expanding ideas, organizing and elaborating sentences, and revising and editing their compositions (Graham, Harris, & Larsen, 2001; Myles, 2002). Computer graphic organizers offer a possible solution to assist them in their writing. This study investigated the effects of a computer graphic organizer on the persuasive writing compositions of Hispanic middle school students with SLD. A multiple baseline design across subjects was used to examine its effects on six dependent variables: number of arguments and supporting details, number and percentage of transferred arguments and supporting details, planning time, writing fluency, syntactical maturity (measured by T-units, the shortest grammatical sentence without fragments), and overall organization. Data were collected and analyzed throughout baseline and intervention. Participants were taught persuasive writing and the writing process prior to baseline. During baseline, participants were given a prompt and asked to use paper and pencil to plan their compositions. A computer was used for typing and editing. Intervention required participants to use a computer graphic organizer for planning and then a computer for typing and editing. The planning sheets and written composition were printed and analyzed daily along with the time each participant spent on planning. The use of computer graphic organizers had a positive effect on the planning and persuasive writing compositions. Increases were noted in the number of supporting details planned, percentage of supporting details transferred, planning time, writing fluency, syntactical maturity in number of T-units, and overall organization of the composition. Minimal to negligible increases were noted in the mean number of arguments planned and written. Varying effects were noted in the percent of transferred arguments and there was a decrease in the T-unit mean length. This study extends the limited literature on the effects of computer graphic organizers as a prewriting strategy for Hispanic students with SLD. In order to fully gauge the potential of this intervention, future research should investigate the use of different features of computer graphic organizer programs, its effects with other writing genres, and different populations.

The Use of a Computer Graphic Organizer for Persuasive Composition Writing by Hispanic Students with Specific Learning Disabilities

Many culturally and linguistically diverse (CLD) students with specific learning disabilities (SLD) struggle with the writing process. Particularly, they have difficulties developing and expanding ideas, organizing and elaborating sentences, and revising and editing their compositions (Graham, Harris, & Larsen, 2001; Myles, 2002). Computer graphic organizers offer a possible solution to assist them in their writing. This study investigated the effects of a computer graphic organizer on the persuasive writing compositions of Hispanic middle school students with SLD. A multiple baseline design across subjects was used to examine its effects on six dependent variables: number of arguments and supporting details, number and percentage of transferred arguments and supporting details, planning time, writing fluency, syntactical maturity (measured by T-units, the shortest grammatical sentence without fragments), and overall organization. Data were collected and analyzed throughout baseline and intervention. Participants were taught persuasive writing and the writing process prior to baseline. During baseline, participants were given a prompt and asked to use paper and pencil to plan their compositions. A computer was used for typing and editing. Intervention required participants to use a computer graphic organizer for planning and then a computer for typing and editing. The planning sheets and written composition were printed and analyzed daily along with the time each participant spent on planning. The use of computer graphic organizers had a positive effect on the planning and persuasive writing compositions. Increases were noted in the number of supporting details planned, percentage of supporting details transferred, planning time, writing fluency, syntactical maturity in number of T-units, and overall organization of the composition. Minimal to negligible increases were noted in the mean number of arguments planned and written. Varying effects were noted in the percent of transferred arguments and there was a decrease in the T-unit mean length. This study extends the limited literature on the effects of computer graphic organizers as a prewriting strategy for Hispanic students with SLD. In order to fully gauge the potential of this intervention, future research should investigate the use of different features of computer graphic organizer programs, its effects with other writing genres, and different populations.

Survivor of Triangle fire and former union organizer

General note: Title and date provided by Bettye Lane.

From the editor

This special issue of Project Management Journal presents a collection of six of the best papers presented at the International Research Network on Organizing by Projects (IRNOP) Conference hosted by the University of Quebec at Montreal (UQAM) in June 2011 in Montréal, Canada. In this editorial, Professor Brian Hobbs, organizer of the IRNOP 2011 Conference and Project Management Chair at UQAM, shares his observations about the current state of project research and identifies trends...

Third International Conference on plant-based vaccines and antibodies

This relatively new biennial meeting - the first was in Prague in 2005 - was chaired by Julian Ma (Guy's Hospital, London, UK), with Mario Pezzotti (University of Verona, Italy) as local organizer, and attracted approximately 180 delegates from 25 countries. The theme was 'Plant Expression Systems for Recombinant Pharmacologics': there were 46 talks gathered into two plenaries, 12 themed sessions and 72 posters. Topics covered included publicly funded and commercial developments, innovation, regulation and commercialization, competition with conventional technology, manufacture and new products. © 2009 Expert Reviews Ltd.

Hashtag as hybrid forum: The case of #agchatoz

This chapter imports Michel Callon’s model of the ‘hybrid forum’ (Callon et al, 2009, p. 18) into social media research, arguing that certain kinds of hashtag publics can be mapped onto this model. It explores this idea of the hashtag as hybrid forum through the worked example of #agchatoz—a hashtag used as both ‘meetup’ organizer for Australian farmers and other stakeholders in Australian agriculture, and as a topic marker for general discussion of related issues. Applying the principles and techniques of digital methods (Rogers, 2013), we employ a standard suite of analytics to a longitudinal dataset of #agchatoz tweets. The results are used not only to describe various elements and dynamics of this hashtag, but also to experiment with the articulation of such approaches with the theoretical model of the hybrid forum, as well as exploring the ways that controversies animate and transform such forums as part of the emergence and cross-pollination of issue publics.