Integrated care and the working record

By default, many discussions and specifications of electronic health records or integrated care records often conceptualize the record as a passive information repository. This article presents data from a case study of work in a medical unit in a major metropolitan hospital. It shows how the clinicians tailored, re-presented and augmented clinical information to support their own roles in the delivery of care for individual patients. This is referred to as the working record: a set of complexly interrelated clinician-centred documents that are locally evolved, maintained and used to support delivery of care in conjunction with the more patient-centred chart that will be stored in the medical records department on the patient’s discharge. Implications are drawn for how an integrated care record could support the local tailorability and flexibility that underpin this working record and hence underpin practice

RNF166 Determines Recruitment of Adaptor Proteins during Antibacterial Autophagy

Xenophagy is a form of selective autophagy that involves the targeting and elimination of intracellular pathogens through several recognition, recruitment, and ubiquitination events. E3 ubiquitin ligases control substrate selectivity in the ubiquitination cascade; however, systematic approaches to map the role of E3 ligases in antibacterial autophagy have been lacking. We screened more than 600 putative human E3 ligases, identifying E3 ligases that are required for adaptor protein recruitment and LC3-bacteria colocalization, critical steps in antibacterial autophagy. An unbiased informatics approach pinpointed RNF166 as a key gene that interacts with the autophagy network and controls the recruitment of ubiquitin as well as the autophagy adaptors p62 and NDP52 to bacteria. Mechanistic studies demonstrated that RNF166 catalyzes K29- and K33-linked polyubiquitination of p62 at residues K91 and K189. Thus, our study expands the catalog of E3 ligases that mediate antibacterial autophagy and identifies a critical role for RNF166 in this process.Leona M. and Harry B. Helmsley Charitable Trust (2014PG-IBD016)National Institutes of Health (U.S.) (R01DK097485)National Institutes of Health (U.S.) (U19AI109725)National Institutes of Health (U.S.) (P30DK043351

Regulation of RNA-dependent RNA polymerase 1 and isochorismate synthase gene expression in Arabidopsis

RNA-dependent RNA polymerases (RDRs) function in anti-viral silencing in Arabidopsis thaliana and other plants. Salicylic acid (SA), an important defensive signal, increases RDR1 gene expression, suggesting that RDR1 contributes to SA-induced virus resistance. In Nicotiana attenuata RDR1 also regulates plant-insect interactions and is induced by another important signal, jasmonic acid (JA). Despite its importance in defense RDR1 regulation has not been investigated in detail.In Arabidopsis, SA-induced RDR1 expression was dependent on 'NON-EXPRESSER OF PATHOGENESIS-RELATED GENES 1', indicating regulation involves the same mechanism controlling many other SA- defense-related genes, including pathogenesis-related 1 (PR1). Isochorismate synthase 1 (ICS1) is required for SA biosynthesis. In defensive signal transduction RDR1 lies downstream of ICS1. However, supplying exogenous SA to ics1-mutant plants did not induce RDR1 or PR1 expression to the same extent as seen in wild type plants. Analysing ICS1 gene expression using transgenic plants expressing ICS1 promoter:reporter gene (β-glucuronidase) constructs and by measuring steady-state ICS1 transcript levels showed that SA positively regulates ICS1. In contrast, ICS2, which is expressed at lower levels than ICS1, is unaffected by SA. The wound-response hormone JA affects expression of Arabidopsis RDR1 but jasmonate-induced expression is independent of CORONATINE-INSENSITIVE 1, which conditions expression of many other JA-responsive genes. Transiently increased RDR1 expression following tobacco mosaic virus inoculation was due to wounding and was not a direct effect of infection. RDR1 gene expression was induced by ethylene and by abscisic acid (an important regulator of drought resistance). However, rdr1-mutant plants showed normal responses to drought.RDR1 is regulated by a much broader range of phytohormones than previously thought, indicating that it plays roles beyond those already suggested in virus resistance and plant-insect interactions. SA positively regulates ICS1

Ethylene and ABA induce <i>AtRDR1</i> expression but RDR1 is not required for drought resistance.

<p>RTqPCR analysis of expression of <i>AtRDR1</i> and the ethylene-regulated gene <i>AtPR4</i> expression in <i>Arabidopsis thaliana</i> Col-0 plants sprayed with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) at 1 mM or water (Control) (A), or of <i>AtRDR1</i> and the ABA-inducible gene <i>RD29A</i> after treatment with ABA (B) over time courses of 72 h. (C) Analysis of water content in wild-type (WT) or <i>rdr1</i>-mutant <i>Arabidopsis</i> plants watered normally (Watered) or deprived of water for 9 days (Drought). In the experiment shown, 40 well-watered plants were divided into two groups of 20, one group watered, the other subjected to drought. There was no significant difference (t-test: <i>p</i> = 0.693) in water content of water-deprived WT and <i>rdr1</i>-transgenic plants. Error bars represent standard error of the mean.</p

Wounding and mock-inoculation induces <i>AtRDR1</i> expression.

<p>(A) <i>AtRDR1</i> expression in TMV-infected, mock-inoculated (Mock) and untreated control (no treatment) <i>Arabidopsis</i> (Col-0) plants over the course of 144 h monitored by RTqPCR (Upper panel). Lower panel shows confirmation by RT-PCR of infection in TMV-inoculated compared with mock-inoculated and untreated (NT, no treatment). (B) RTqPCR of expression of <i>AtRDR1</i> and the wounding- and JA-responsive gene <i>AtTPS10</i> over 72 h following wounding. Error bars represent standard error of the mean.</p

SA induces <i>AtRDR1</i> expression in an NPR1-dependent manner.

<p>(A) <i>AtRDR1</i> and <i>AtPR1</i> expression in ecotype Col-0 wild type and <i>npr1-1</i> control and SA treated plants 6 h after treatment. (B) <i>AtRDR1</i> and <i>AtPR1</i> expression in Nössen (NO) wild-type and <i>npr1-5</i> control and SA treated plants 6 h after treatment. Error bars represent standard error of the mean.</p

<i>AtICS1</i> expression is positively regulated by SA and is required for optimal expression of <i>AtRDR1</i> and <i>PR1</i> in response to exogenous SA treatment.

<p>(A) RTqPCR analysis of transcript accumulation for <i>AtRDR1</i> and <i>AtPR1</i> expression in <i>Arabidopsis</i> ecotype Col-0 wild type (WT), <i>sid2</i>-mutant (compromised in expression of <i>ICS1</i>) and <i>NahG</i>-transgenic plants 6 h post SA treatment. (B) Transgenic <i>Arabidopsis</i> plants harboring the promoter: reporter constructs <i>ICS1:GUS</i> and <i>ICS2:GUS</i> stained for GUS activity 24 h after control (water) or SA treatment. SA treated Col-0 WT and <i>35S:GUS</i> plants, 24 h after treatment are included as controls. (C) RTqPCR analysis of <i>AtICS1</i>, <i>AtICS2</i>, and <i>AtPR1</i> transcript accumulation in wild-type plants treated with SA, over a 48 h time course. Error bars represent standard error of the mean.</p

JA-induced <i>AtRDR1</i> expression is COI1-independent.

<p>(A) RTqPCR analysis of transcript accumulation for <i>AtRDR1</i> and the JA-responsive, COI1-dependent gene <i>AtTPS10</i> over 72 h following treatment of <i>Arabidopsis</i> (ecotype Col-0) plants with methyl-JA. (B) <i>AtRDR1</i> and <i>AtTPS10</i> transcript accumulation in methyl-JA (JA) or control-treated <i>coi1-16</i> mutant plants and wild-type Col-0 or Col-0 gl (the <i>coi1-16</i> background) at 6 h post-treatment. Error bars represent standard error of the mean.</p