Material and Process Parameters that Affect Accuracy in Stereolithography

Experimental real time linear shrinkage rate measurements simulating stereolithography are used in an analysis of shrinkage during line drawing in stereolithography. While the amount of shrinkage depends on the polymerization kinetics, shrinkage kinetics and overall degree of cure, it also depends on the length of time to draw a line of plastic. A line drawn slowly will exhibit less apparent shrinkage than one drawn very quickly because much of the shrinkage is compensated for as the line is drawn. The data also indicates that a typical stereolithography resin in the green state may shrink to only 65% of its maximum, thus retaining considerable potential for shrinkage during post-cure. This infonnation can be used to predict the amount of shrinkage to be expected under certain exposure conditions and to fonnulate overall strategies to reduce shrinkage and subsequent warpage that causes shape distortion.Mechanical Engineerin

Saturn Forms by Core Accretion in 3.4 Myr

We present two new in situ core accretion simulations of Saturn with planet formation timescales of 3.37 Myr (model S0) and 3.48 Myr (model S1), consistent with observed protostellar disk lifetimes. In model S0, we assume rapid grain settling reduces opacity due to grains from full interstellar values (Podolak 2003). In model S1, we do not invoke grain settling, instead assigning full interstellar opacities to grains in the envelope. Surprisingly, the two models produce nearly identical formation timescales and core/atmosphere mass ratios. We therefore observe a new manifestation of core accretion theory: at large heliocentric distances, the solid core growth rate (limited by Keplerian orbital velocity) controls the planet formation timescale. We argue that this paradigm should apply to Uranus and Neptune as well.Comment: 4 pages, including 1 figure, submitted to ApJ Letter

Mechanism of Cross-Species Prion Transmission An Infectious Conformation Compatible withTwo Highly Divergent Yeast Prion Proteins

SummaryEfficiency of interspecies prion transmission decreases as the primary structures of the infectious proteins diverge. Yet, a single prion protein can misfold into multiple infectious conformations, and such differences in “strain conformation” also alter infection specificity. Here, we explored the relationship between prion strains and species barriers by creating distinct synthetic prion forms of the yeast prion protein Sup35. We identified a strain conformation of Sup35 that allows transmission from the S. cerevisiae (Sc) Sup35 to the highly divergent C. albicans (Ca) Sup35 both in vivo and in vitro. Remarkably, cross-species transmission leads to a novel Ca strain that in turn can infect the Sc protein. Structural studies reveal strain-specific conformational differences in regions of the prion domain that are involved in intermolecular contacts. Our findings support a model whereby strain conformation is the critical determinant of cross-species prion transmission while primary structure affects transmission specificity by altering the spectrum of preferred amyloid conformations

Expression of TCR-Vβ peptides by murine bone marrow cells does not identify T-cell progenitors

Germline transcription has been described for both immunoglobulin and T-cell receptor (TCR) genes, raising questions of their functional significance during haematopoiesis. Previously, an immature murine T-cell line was shown to bind antibody to TCR-Vβ8.2 in absence of anti-Cβ antibody binding, and an equivalent cell subset was also identified in the mesenteric lymph node. Here, we investigate whether germline transcription and cell surface Vβ8.2 expression could therefore represent a potential marker of T-cell progenitors. Cells with the TCR phenotype of Vβ8.2(+) Cβ(-) are found in several lymphoid sites, and among the lineage-negative (Lin(-) ) fraction of hematopoietic progenitors in bone marrow (BM). Cell surface marker analysis of these cells identified subsets reflecting common lymphoid progenitors, common myeloid progenitors and multipotential progenitors. To assess whether the Lin(-) Vβ8.2(+) Cβ(-) BM subset contains hematopoietic progenitors, cells were sorted and adoptively transferred into sub-lethally irradiated recipients. No T-cell or myeloid progeny were detected following introduction of cells via the intrathymic or intravenous routes. However, B-cell development was detected in spleen. This pattern of restricted in vivo reconstitution disputes Lin(-) Vβ8.2(+) Cβ(-) BM cells as committed T-cell progenitors, but raises the possibility of progenitors with potential for B-cell development.This study was supported by grants to H.C.O. from the National Health and Medical Research Council of Australia. J.A. was supported by a fellowship from the Australian Academy of Science

Developing a collaborative, humanistic interprofessional healthcare culture: a multi-site study

Introduction:Developing a collaborative, humanistic interprofessional healthcare culture requires optimal relational skills, respect, interpersonal cohesion, and role clarity. We developed a longitudinal curriculum to engender these skills and values in institutional leaders. We report results of a qualitative study at seven US-based academic health centers to identify participants’ learning. Methods:At each institution, participants from at least three different professions met in small group sessions twice-monthly over nine months. Sessions focused on relational capacities to enhance leadership and professionalism, and utilized critical reflection and experiential learning to promote teamwork, self-knowledge, communication skills, and address challenges encountered by a healthcare team. Participants completed reflective responses to open-ended questions asking what knowledge, insights, or skills they gained by working in this interprofessional group and applications of their learning. Five investigators analyzed the anonymized responses using the constant comparative method. Results:Overarching themes centered on relationships and the strength of the relational nature of the learning. We observed learning on three levels: a) Intrapersonal learning included self-awareness, mindfulness, and empathy for self that translated to reflections on application of these to teamwork and teaching; b) Interpersonal learning concerned relational skills and teaching about listening, understanding others’ perspectives, appreciation/respect for colleagues, and empathy for others; c) Systems level learning included teaching skills about resilience, conflict management, team dynamics and cultural norms, and appreciation of resources from interprofessional colleagues. Discussion:A curriculum focusing on humanistic teaching for leaders led to new insights and positive changes in relational perspectives. Learning occurred on multiple levels. Many learners reported revising previous assumptions, a marker for transformative learning. Humanistic faculty development can facilitate deep bonds between professions. &nbsp

The helicase Ded1p controls use of near-cognate translation initiation codons in 5' UTRs.

The conserved and essential DEAD-box RNA helicase Ded1p from yeast and its mammalian orthologue DDX3 are critical for the initiation of translation1. Mutations in DDX3 are linked to tumorigenesis2-4 and intellectual disability5, and the enzyme is targeted by a range of viruses6. How Ded1p and its orthologues engage RNAs during the initiation of translation is unknown. Here we show, by integrating transcriptome-wide analyses of translation, RNA structure and Ded1p-RNA binding, that the effects of Ded1p on the initiation of translation are connected to near-cognate initiation codons in 5' untranslated regions. Ded1p associates with the translation pre-initiation complex at the mRNA entry channel and repressing the activity of Ded1p leads to the accumulation of RNA structure in 5' untranslated regions, the initiation of translation from near-cognate start codons immediately upstream of these structures and decreased protein synthesis from the corresponding main open reading frames. The data reveal a program for the regulation of translation that links Ded1p, the activation of near-cognate start codons and mRNA structure. This program has a role in meiosis, in which a marked decrease in the levels of Ded1p is accompanied by the activation of the alternative translation initiation sites that are seen when the activity of Ded1p is repressed. Our observations indicate that Ded1p affects translation initiation by controlling the use of near-cognate initiation codons that are proximal to mRNA structure in 5' untranslated regions

Versatile in vivo regulation of tumor phenotypes by dCas9-mediated transcriptional perturbation

Targeted transcriptional regulation is a powerful tool to study genetic mediators of cellular behavior. Here, we show that catalytically dead Cas9 (dCas9) targeted to genomic regions upstream or downstream of the transcription start site allows for specific and sustainable gene-expression level alterations in tumor cells in vitro and in syngeneic immune-competent mouse models. We used this approach for a high-coverage pooled gene-activation screen in vivo and discovered previously unidentified modulators of tumor growth and therapeutic response. Moreover, by using dCas9 linked to an activation domain, we can either enhance or suppress target gene expression simply by changing the genetic location of dCas9 binding relative to the transcription start site. We demonstrate that these directed changes in gene-transcription levels occur with minimal off-target effects. Our findings highlight the use of dCas9-mediated transcriptional regulation as a versatile tool to reproducibly interrogate tumor phenotypes in vivo. Keywords: cancer genetics; cancer models; cancer therapeutic resistance; gene regulation; CRISPRNational Cancer Institute (U.S.) (Grant U54-CA112967-06)National Cancer Institute (U.S.) (Grant P30-CA14051