2,600 research outputs found

    The Hippocampus as a Cognitive Map

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    Ectopic cyclin E expression induces premature entry into S phase and disrupts pattern formation in the Drosophila eye imaginal disc

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    During animal development, cell proliferation is controlled in many cases by regulation of the G1 to S phase transition. Studies of mammalian tissue culture cells have shown that the G1-specific cyclin, cyclin E, can be rate limiting for progression from G1 to S phase. During Drosophila development, down-regulation of cyclin E is required for G1 arrest in terminally differentiating embryonic epidermal cells. Whether cyclin E expression limits progression into S phase in proliferating, as opposed to differentiating, cells during development has not been investigated. Here we show that Drosophila cyclin E (DmcycE) protein is absent in G1 phase cells but appears at the onset of S phase in proliferating cells of the larval optic lobe and eye imaginal disc. We have examined cells in the eye imaginal epithelium, where a clearly defined developmentally regulated G1 to S phase transition occurs. Ectopic expression of DmcycE induces premature entry of most of these G1 cells into S phase. Thus in these cells, control of DmcycE expression is required for regulated entry into S phase. Significantly, a band of eye imaginal disc cells in G1 phase was not induced to enter S phase by ectopic expression of DmcycE. This provides evidence for additional regulatory mechanisms that operate during G1 phase to limit cell proliferation during development. These results demonstrate that the role of cyclin E in regulating progression into S phase in mammalian tissue culture cells applies to some, but not all, cells during Drosophila development. Ectopic expression of DmcycE in the eye imaginal disc disrupts normal pattern formation, highlighting the importance of coordinating cell proliferation with developmental processes for correct patterning in the developing eye. These studies establish DmcycE as a target of regulatory mechanisms that coordinate cell proliferation with other developmental events

    Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences

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    Five distinct patterns of DNA replication have been identified during S-phase in asynchronous and synchronous cultures of mammalian cells by conventional fluorescence microscopy, confocal laser scanning microscopy, and immunoelectron microscopy. During early S-phase, replicating DNA (as identified by 5-bromodeoxyuridine incorporation) appears to be distributed at sites throughout the nucleoplasm, excluding the nucleolus. In CHO cells, this pattern of replication peaks at 30 min into S-phase and is consistent with the localization of euchromatin. As S-phase continues, replication of euchromatin decreases and the peripheral regions of heterochromatin begin to replicate. This pattern of replication peaks at 2 h into S-phase. At 5 h, perinucleolar chromatin as well as peripheral areas of heterochromatin peak in replication. 7 h into S-phase interconnecting patches of electron-dense chromatin replicate. At the end of S-phase (9 h), replication occurs at a few large regions of electron-dense chromatin. Similar or identical patterns have been identified in a variety of mammalian cell types. The replication of specific chromosomal regions within the context of the BrdU-labeling patterns has been examined on an hourly basis in synchronized HeLa cells. Double labeling of DNA replication sites and chromosome-specific alpha-satellite DNA sequences indicates that the alpha-satellite DNA replicates during mid S-phase (characterized by the third pattern of replication) in a variety of human cell types. Our data demonstrates that specific DNA sequences replicate at spatially and temporally defined points during the cell cycle and supports a spatially dynamic model of DNA replication

    Spitz and wingless, emanating from distinct borders, cooperate to establish cell fate across the engrailed domain in the drosophila epidermis

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    A key step in development is the establishment of cell type diversity across a cellular field. Segmental patterning within the Drosophila embryonic epidermis is one paradigm for this process. At each parasegment boundary, cells expressing the Wnt family member Wingless confront cells expressing the homeoprotein Engrailed. The Engrailed-expressing cells normally differentiate as one of two alternative cell types. In investigating the generation of this cell type diversity among the 2-cell-wide Engrailed stripe, we previously showed that Wingless, expressed just anterior to the Engrailed cells, is essential for the specification of anterior Engrailed cell fate. In a screen for additional mutations affecting Engrailed cell fate, we identified anterior open/yan, a gene encoding an inhibitory ETS-domain transcription factor that is negatively regulated by the Rasl-MAP kinase signaling cascade. We find that Anterior Open must be inactivated for posterior Engrailed cells to adopt their correct fate. This is achieved by the EGF receptor (DER), which is required autonomously in the Engrailed cells to trigger the Ras1-MAP kinase pathway. Localized activation of DER is accomplished by restricted processing of the activating ligand, Spitz. Processing is confined to the cell row posterior to the Engrailed domain by the restricted expression of Rhomboid. These cells also express the inhibitory ligand Argos, which attenuates the activation of DER in cell rows more distant from the ligand source. Thus, distinct signals flank each border of the Engrailed domain, as Wingless is produced anteriorly and Spitz posteriorly. Since we also show that En cells have the capacity to respond to either Wingless or Spitz, these cells must choose their fate depending on the relative level of activation of the two pathways.Louise O’Keefe, Scott T. Dougan, Limor Gabay, Erez Raz, Ben-Zion Shilo and Stephen DiNard

    In-between spaces of policy and practice: Voices from Prince Edward Island early childhood educators

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    Over the course of the past decades, the discourse, pedagogy, scope, and delivery of early learning and child care (ELCC) has undergone myriad significant changes internationally, nationally, and at local levels. Prince Edward Island (PEI), the smallest Canadian Province, has not been exempt from these transformations. By situating early childhood educators (ECEs) at the centre of ecological multilevel environments (Bronfenbrenner, 2005), this qualitative study explored how a system-wide change implemented through the Prince Edward Island Preschool Excellence Initiative (PEIPEI) has impacted and is being impacted by ECEs over time. Purposive sampling was used to invite seven early childhood educators working on provincially regulated early years centres (EYCs) to participate in individual interviews. Findings indicated that ECEs have been striving to navigate and merge the space in-between policy and practices and that after ten years, they remain in this liminal space where they continue to navigate unravelling transitions as they search for their professional identity

    Medium-term follow-up of ulcerative colitis in Cape Town

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    The 114 patients with ulcerative colitis diagnosed in Greater Cape Town between 1970 and 1979 were followed up 11 years later. Ninety per cent of those contacted were in remission or had mild symptoms only. Eleven patients had died; 3 deaths (in total colitis patients) were disease-related but the overall mortality rate in ulcerative colitis was not increased. There was only 1 case of carcinoma of the colon. The 5-year surgical rate was 5% increasing to 23% 10 years after diagnosis. Six patients (35%) had had a Park's pouch, 3 (18%) ileorectal anastomosis, and a (47%) panproctocolectomy or colectomy with an ileostomy. The incidence of surgery was higher in those with total colitis. In those patients who did not have the rectum removed, there was a 100% recurrence of proctitis. Park's pouch patients remained well and incontinence was not a problem. Thirty-one per cent of patients with proctitis at diagnosis had evidence of extension of disease to the colon at follow-up. Ulcerative colitis may be a more benign disease than often believed, with mortality from the disease and need for surgery being associated almost exclusively with extensive disease.S Afr Med J 1989; 76: 142·14

    Disruption of pre-mRNA splicing in vivo results in reorganization of splicing factors

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    We have examined the functional significance of the organization of pre-mRNA splicing factors in a speckled distribution in the mammalian cell nucleus. Upon microinjection into living cells of oligonucleotides or antibodies that inhibit pre-mRNA splicing in vitro, we observed major changes in the organization of splicing factors in vivo. Interchromatin granule clusters became uniform in shape, decreased in number, and increased in both size and content of splicing factors, as measured by immunofluorescence. These changes were transient and the organization of splicing factors returned to their normal distribution by 24 h following microinjection. Microinjection of these oligonucleotides or antibodies also resulted in a reduction of transcription in vivo, but the oligonucleotides did not inhibit transcription in vitro. Control oligonucleotides did not disrupt splicing or transcription in vivo. We propose that the reorganization of splicing factors we observed is the result of the inhibition of splicing in vivo

    Consumer responses to a future UK food system

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    Purpose– The purpose of this paper is to describe research exploring consumer responses to potential changes in food-related practices to mitigate and adapt to climate change.Design/methodology/approach– Six focus groups explored consumer responses to measures to intended to mitigate the emissions from, and adapt to the impacts of climate change. These included: meat reduction, greater reliance on seasonal British food, meal replacement tablets, laboratory grown meat, communal eating houses, genetically modified food and food waste. Practice theory provided the lens to interpret the changes to meanings, competences and materials associated with food consumption.Findings– Changes that could be assimilated within existing competencies were viewed more positively, with lack of competence a key barrier to accommodating change. At present, climate change and sustainability do not influence purchasing decisions. Policy measures delivering multiple benefits (“win-wins”), of which environmental performance may be one, stand an improved chance of establishing more sustainable practices than those focusing exclusively on environmental drivers.Originality/value– Awareness of the role of sustainable food systems in the context of anthropogenic climate change is growing. Whilst scientific and technological research explores methods for reducing emissions and building resilience in food supply chains to changes in climate, there is comparatively little study of how consumers perceive these proposed “solutions”. This research provides a comprehensive overview of consumer responses to potential changes in eating practices related to climate change mitigation and adaptation and is of value to policy makers, academics and practitioners across the food supply chain.</jats:sec
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