Homozygous knockout of the piezo1 gene in the zebrafish is not associated with anemia

We have now examined the erythroid phenotype in this zebrafish strain carrying a ZFN genomic knockout of piezo1. Genotyping was performed as previously described. In contrast to the anemic phenotype observed in zebrafish subjected to morpholino knockdown of piezo, the genomic ZFN knockout of piezo1 did not segregate either with anemia in the 3-dpf embryo or with dysmorphic erythrocyte morphology in the adult fish

5,6-Di­oxo-1,10-phenanthrolin-1-ium trifluoro­methane­sulfonate

In the structure of the title salt, C12H7N2O2 +·CF3SO3 −, the cation participates in hydrogen bonding with the dione group of an adjacent cation as well as with the trifluoro­methane­sulfonate anion. In addition, there is an extensive network of C—H⋯O inter­actions between the cations and anions. There are two formula units per asymmetric unit. The crystal studied exhibits inversion twinning

Acetato(1,10-phenanthroline-5,6-dione)silver(I) trihydrate

In the structure of the title compound, [Ag(C2H3O2)(C12H6N2O2)]·3H2O, the AgI atom is coordinated by both 1,10-phenanthroline-5,6-dione N atoms and one O atom from the acetate anion. The three water mol­ecules are involved in extensive hydrogen bonding to each other and to the acetate O and 1,10-phenanthroline-5,6-dione O atoms. In addition, there are weak C—H⋯O inter­actions

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

Split-wrmScarlet and split-sfGFP: tools for faster, easier fluorescent labeling of endogenous proteins in Caenorhabditis elegans

&lt;p&gt;Zenodo hosts the archival version of this document; for convenient viewing, please visit&nbsp;&lt;a href="http://andrewgyork.github.io/split_wrmscarlet" target="_blank" rel="noopener"&gt;andrewgyork.github.io/split_wrmscarlet &lt;/a&gt;or&nbsp;&lt;a href="http://marimar128.github.io/split_wrmscarlet" target="_blank" rel="noopener"&gt;marimar128.github.io/split_wrmscarlet&lt;/a&gt;.&lt;/p&gt; &lt;h4&gt;&lt;a href="https://andrewgyork.github.io/split_wrmscarlet/#Abstract" target="_self"&gt;Abstract&lt;/a&gt;&lt;/h4&gt; &lt;p&gt;We create and share a new red fluorophore, along with a set of strains, reagents and protocols, to make it faster and easier to label endogenous&nbsp;&lt;em&gt;C. elegans&lt;/em&gt;&nbsp;proteins with fluorescent tags. CRISPR-mediated fluorescent labeling of&nbsp;&lt;em&gt;C. elegans&lt;/em&gt;&nbsp;proteins is an invaluable tool, but it is much more difficult to insert fluorophore-size DNA segments than it is to make small gene edits. In principle, high-affinity asymmetrically-split fluorescent proteins solve this problem in&nbsp;&lt;em&gt;C. elegans&lt;/em&gt;: the small fragment can quickly and easily be fused to almost any protein of interest and can be detected wherever the large fragment is expressed and complemented. However, there is currently only one available strain stably expressing the large fragment of a split fluorescent protein, restricting this solution to a single tissue (the germline) in the highly autofluorescent green channel. No available&nbsp;&lt;em&gt;C. elegans&lt;/em&gt;&nbsp;lines express unbound large fragments of split red fluorescent proteins, and even state-of-the-art split red fluorescent proteins are dim compared to the canonical split-sfGFP protein. In this study, we engineer a bright, high-affinity new split red fluorophore,&nbsp;&lt;a href="https://www.addgene.org/138966/"&gt;split-wrmScarlet&lt;/a&gt;. We generate transgenic&nbsp;&lt;em&gt;C. elegans&lt;/em&gt;&nbsp;lines to allow easy&nbsp;&lt;a href="https://cgc.umn.edu/strain/CF4582"&gt;single-color&lt;/a&gt;&nbsp;labeling in&nbsp;&lt;a href="https://cgc.umn.edu/strain/CF4610"&gt;muscle&lt;/a&gt;&nbsp;or&nbsp;&lt;a href="https://cgc.umn.edu/strain/DUP237"&gt;germline&lt;/a&gt;&nbsp;and&nbsp;&lt;a href="https://cgc.umn.edu/strain/CF4588"&gt;dual-color&lt;/a&gt;&nbsp;labeling in somatic cells. We also describe 'glonads', a novel expression strategy for the germline, where traditional expression strategies struggle. We validate these strains by targeting split-wrmScarlet to several genes whose products label distinct organelles, and we provide a&nbsp;&lt;a href="https://doi.org/10.17504/protocols.io.bamkic4w"&gt;protocol&lt;/a&gt;&nbsp;for easy, cloning-free CRISPR/Cas9 editing. As the collection of split-FP strains for labeling in different tissues or organelles expands, we will post updates at&nbsp;&lt;a href="https://doi.org/10.5281/zenodo.3993663"&gt;doi.org/10.5281/zenodo.3993663&lt;/a&gt;&lt;/p&gt

Cutting Edge: Self-Antigen Controls the Balance between Effector and Regulatory T Cells in Peripheral Tissues

Immune homeostasis in peripheral tissues is achieved by maintaining a balance between pathogenic effector T cells (Teff) and protective Foxp3(+) regulatory T cells (Treg). Using a mouse model of an inducible tissue-antigen we demonstrate that antigen (Ag) persistence is a major determinant of the relative frequencies of Teff and Treg cells. Encounter of transferred naïve CD4(+) T cells with transiently expressed tissue-Ag leads to generation of cytokine-producing Teff cells and peripheral Treg cells. Persistent expression of Ag, a mimic of self Ag, leads to functional inactivation and loss of the Teff cells with preservation of Treg in the target tissue. The inactivation of Teff cells by persistent Ag is associated with reduced ERK phosphorylation (pERK), whereas Treg cells show less reduction in pERK and are relatively resistant to ERK inhibition. Our studies reveal a crucial role for Ag in maintaining appropriate ratios of Ag-specific Teff to Treg cells in tissues