Discerning lines of demarcation

Discerning Lines of Demarcation is an investigation into the accumulated landscape of distressing times. Situations of mass destruction, loss of family, substance abuse, domestic violence, loss of friendship, and uncertain health have all been encountered within a steady progression in the last five to six years of my life. The digestion of these situations has been slow as the events overlap and intertwine each other. I have tweezed and distilled these circumstances. This is described through different types of terrains that create physical boundaries to represent psychological fears or events. Tied to Your Hate; How Much More Will Fall, Untitled, Loss Can Ruin but Growth Can Be Found, and Precarious Footing are all pieces that reach for a greater understanding of many trying events. The work uses the format of installation as the vehicle in which to articulate the landscapes or yards. This format allows the viewer to be inside the workings of the various pieces

Harris v. Quinn and the Contradictions of Compelled Speech

Krpelji su parazitski člankonošci iz reda grinja koji je dio razreda paučnjaka. Dijele se na tvrde krpelje Ixodidae (krpelji šikare; 692 vrste), meke krpelje Argasidae (krpelje nastambe; 186 vrsta) i Nuttalliellidae koja sadržava samo jednu vrstu. Uzorkovanje tvrdih krpelja (Acari: Ixodidae) na području Rekreacijsko športskog centra Jarun obavljeno je od ožujka do rujna 2018. godine. Ukupno je prikupljeno 88 jedinki. Determinacijom je ustanovljeno da sve jedinke pripadaju istoj vrsti - obični ili šumski krpelj (Ixodes ricinus). Krpelji su lovljeni metodom krpeljne zatege na pet različitih mikrolokaliteta. Među uzorkovanim krpeljima utvrđena je dominacija krpelja u razvojnom stadiju larve. Najveći broj uzorkovanih jedinki vrste Ixodes ricinus bio je u srpnju, dok je najmanji bio u kolovozu. Krpelji su glavni prijenosnici zoonoza (lajmske borelioze, krpeljnog meningoencefalitisa, tularemije) kod ljudi i životinja, te je stoga bitno utvrditi njihovu brojnost, te educirati javnost kako bi se podigla svijest o realnoj opasnosti od ugriza krpelja

Harris v. Quinn and the Contradictions of Compelled Speech

In Harris v. Quinn, the Supreme Court held that unionized homecare workers have a First Amendment right to refuse to pay their fair share of the cost of services that the union is statutorily required to provide. The Court thus transformed what had been a legislative debate about “right-to-work” laws, which about half of states have adopted, into a constitutional requirement for one narrow category of public sector employees. The problem with transforming this policy argument into a First Amendment requirement is that treating fair-share or agency-fee payments to a union as compelled speech raises First Amendment rights of both supporters and opponents of the union. If expenditures on union representation are speech—as the majority in Harris thinks they are—then the union’s obligation to provide free representation compels speech by the union and its members. While, in our view, the requirement to pay for services is not compelled speech, the Court’s entire agency-fee jurisprudence, including Harris, insists that it is. On the Court’s analysis, contracts that require unionized employees to pay for union representational services compel speech of dissenters exactly to the same extent that their prohibition compels speech of unions and their members. Accordingly, the Court must alter its usual analysis of the constitutionality of agency-fee agreements and recognize that union representation requires balancing competing freedom of speech and association interests. Once the First Amendment rights of unions and union members are recognized, agency fees emerge as a constitutionally sound accommodation of the interests of dissenters, unions, and union members

Rapid validation of cancer genes in chimeras derived from established genetically engineered mouse models

Recent technological advances have opened the door for the fast and cost-effective generation of genetically engineered mouse models (GEMMs) to study cancer. We describe here a conceptually novel approach for the generation of chimeric GEMMs based on the controlled introduction of various genetic elements in embryonic stem cells (ESCs) that are derived from existing mouse strains with a predisposition for cancer. The isolation of GEMM-derived ESC lines is greatly facilitated by the availability of the newly defined culture media containing inhibitors that effectively preserve ESC pluripotency. The feasibility of the GEMM-ESC approach is discussed in light of current literature and placed into the context of existing models. This approach will allow for fast and flexible validation of candidate cancer genes and drug targets and will result in a repository of GEMM-ESC lines and corresponding vector collections that enable easy distribution and use of preclinical models to the wider scientific community

Multiple knockout mouse models reveal lincRNAs are required for life and brain development

Many studies are uncovering functional roles for long noncoding RNAs (lncRNAs), yet few have been tested for in vivo relevance through genetic ablation in animal models. To investigate the functional relevance of lncRNAs in various physiological conditions, we have developed a collection of 18 lncRNA knockout strains in which the locus is maintained transcriptionally active. Initial characterization revealed peri- and postnatal lethal phenotypes in three mutant strains (Fendrr, Peril, and Mdgt), the latter two exhibiting incomplete penetrance and growth defects in survivors. We also report growth defects for two additional mutant strains (linc–Brn1b and linc–Pint). Further analysis revealed defects in lung, gastrointestinal tract, and heart in Fendrr−/− neonates, whereas linc–Brn1b−/− mutants displayed distinct abnormalities in the generation of upper layer II–IV neurons in the neocortex. This study demonstrates that lncRNAs play critical roles in vivo and provides a framework and impetus for future larger-scale functional investigation into the roles of lncRNA molecules. DOI: http://dx.doi.org/10.7554/eLife.01749.00

Butyrophilin-like 2 regulates site-specific adaptations of intestinal γδ intraepithelial lymphocytes

Tissue-resident γδ intraepithelial lymphocytes (IELs) orchestrate innate and adaptive immune responses to maintain intestinal epithelial barrier integrity. Epithelia-specific butyrophilin-like (Btnl) molecules induce perinatal development of distinct Vγ TCR+ IELs, however, the mechanisms that control γδ IEL maintenance within discrete intestinal segments are unclear. Here, we show that Btnl2 suppressed homeostatic proliferation of γδ IELs preferentially in the ileum. High throughput transcriptomic characterization of site-specific Btnl2-KO γδ IELs reveals that Btnl2 regulated the antimicrobial response module of ileal γδ IELs. Btnl2 deficiency shapes the TCR specificities and TCRγ/δ repertoire diversity of ileal γδ IELs. During DSS-induced colitis, Btnl2-KO mice exhibit increased inflammation and delayed mucosal repair in the colon. Collectively, these data suggest that Btnl2 fine-tunes γδ IEL frequencies and TCR specificities in response to site-specific homeostatic and inflammatory cues. Hence, Btnl-mediated targeting of γδ IEL development and maintenance may help dissect their immunological functions in intestinal diseases with segment-specific manifestations

Efficient Generation of Germ Line Transmitting Chimeras from C57BL/6N ES Cells by Aggregation with Outbred Host Embryos

Genetically modified mouse strains derived from embryonic stem (ES) cells have become essential tools for functional genomics and biomedical research. Large scale mutagenesis projects are producing libraries of mutant C57BL/6 (B6) ES cells to enable the functional annotation of every gene of the mouse genome. To realize the utility of these resources, efficient and accessible methods of generating mutant mice from these ES cells are necessary. Here, we describe a combination of ICR morula aggregation and a chemically-defined culture medium with widely available and accessible components for the high efficiency generation of germline transmitting chimeras from C57BL/6N ES cells. Together these methods will ease the access of the broader biomedical research community to the publicly available B6 ES cell resources

Marking Embryonic Stem Cells with a 2A Self-Cleaving Peptide: A NKX2-5 Emerald GFP BAC Reporter

Fluorescent reporters are useful for assaying gene expression in living cells and for identifying and isolating pure cell populations from heterogeneous cultures, including embryonic stem (ES) cells. Multiple fluorophores and genetic selection markers exist; however, a system for creating reporter constructs that preserve the regulatory sequences near a gene's native ATG start site has not been widely available.Here, we describe a series of modular marker plasmids containing independent reporter, bacterial selection, and eukaryotic selection components, compatible with both Gateway recombination and lambda prophage bacterial artificial chromosome (BAC) recombineering techniques. A 2A self-cleaving peptide links the reporter to the native open reading frame. We use an emerald GFP marker cassette to create a human BAC reporter and ES cell reporter line for the early cardiac marker NKX2-5. NKX2-5 expression was detected in differentiating mouse ES cells and ES cell-derived mice.Our results describe a NKX2-5 ES cell reporter line for studying early events in cardiomyocyte formation. The results also demonstrate that our modular marker plasmids could be used for generating reporters from unmodified BACs, potentially as part of an ES cell reporter library