Association of pre-operative medication use with post-operative delirium in surgical oncology patients receiving comprehensive geriatric assessment

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.Abstract Background Older patients undergoing surgery tend to have a higher frequency of delirium. Delirium is strongly associated with poor surgical outcomes. This study evaluated the association between pre-operative medication use and post-operative delirium (POD) in surgical oncology patients receiving comprehensive geriatric assessment (CGA). Methods A total of 475 patients who were scheduled for cancer surgery and received CGA from January 2014 to June 2015 were included. Pre-operative medication review through CGA was conducted on polypharmacy (≥5 medications), delirium-inducing medications (DIMs), fall-inducing medications (FIMs), and potentially inappropriate medications (PIMs). POD was confirmed by psychiatric consultation, and DSM-V criteria were used for diagnosing delirium. The model fit of the prediction model was assessed by computing the Hosmer-Lemeshow goodness-of-fit test. Effect size was measured using the Nagelkerke R2. Discrimination of the model was assessed by an analysis of the area under receiver operating curve (AUROC). Results Two models were constructed for multivariate analysis based on univariate analysis; model I included dementia and DIM in addition to age and sex, and model II included PIM instead of DIM of model I. Every one year increase of age increased the risk of POD by about 1.1-fold. DIM was a significant factor for POD after adjusting for confounders (AOR 12.78, 95 % CI 2.83-57.74). PIM was also a significant factor for POD (AOR 5.53, 95 % CI 2.03-15.05). The Hosmer-Lemeshow test results revealed good fits for both models (χ2 = 3.842, p = 0.871 for model I and χ2 = 8.130, p = 0.421 for model II). The Nagelkerke R2 effect size and AUROC for model I was 0.215 and 0.833, respectively. Model II had the Nagelkerke R2effect size of 0.174 and AUROC of 0.819. Conclusions These results suggest that pharmacists comprehensive review for pre-operative medication use is critical for the post-operative outcomes like delirium in older patients

Molecular characterization of stress-inducible PLATZ gene from soybean (Glycine max L.)

Abstract PLATZ (plant AT-rich sequence and zinc-binding protein) is a novel class of DNA-binding proteins; however, the function of the PLATZ gene has not yet been identified in plants. This study aims to isolate, sequence, and analyse the PLATZ gene responsive to abiotic stress in soybean. We isolated a stress-inducible gene encoding the PLATZ from soybean (Glycine max L.). This gene, designated as GmPLATZ1, was specifically induced by drought, high salinity, or abscisic acid (ABA) in soybean. GmPLATZ1 cDNA is composed of 711 bp nucleotide sequences encoding a PLATZ protein (236 amino acids) with 9.41 pI and 26.75 kDa. Multiple sequence alignment analysis showed that the N-terminal region of GmPLATZ1 shares the highly conserved zinc-finger motifs with other PLATZ proteins. The subcellular localization of the GmPLATZ1 protein was analysed via the green fluorescent protein (GFP)-GmPLATZ1 fusion protein in tobacco plant cell. The GFP-GmPLATZ1 protein was shown to be targeted to the nucleus. The function of the GmPLATZ1 gene was further investigated using the overexpression transgenic Arabidopsis. The germination rate in transgenic plants overexpressing GmPLATZ1 was significantly delayed in media supplemented with mannitol compared with that of wild-type (WT) plants. Moreover, cotyledon development in the GmPLATZ1-overexpression transgenic plants was remarkably retarded in the presence of ABA compared to WT. This shows that GmPLATZ1 is implicated in developmental processes such as germination under osmotic stress conditions in plants

Tolerance tests of wild-type and <i>35S:VrUBC1</i> Arabidopsis transgenic plants under osmotic stress or ABA conditions.

<p>(A) Ten-d-old seedlings of the wild-type and <i>35S:VrUBC1</i> Arabidopsis transgenic lines (L19 and L23) were transferred to MS medium containing 2% (w/v) sucrose and 0.8% (w/v) phytoagar supplemented either mannitol (200 mM) or with NaCl (150 mM). (B) Root length was monitored after 10 days. The values are the means ± SD (n = 3). This experiment was carried out three times with consistent results.</p

Interaction between E3 ligases and VrUBC1 in yeast and <i>in vitro</i> self-ubiquitination of AtVBP1.

<p>(A) Protein interactions of VrUBC1 with five C3HC4-type RING E3 ligases and three positive regulators for osmotic tolerance such as RHA2a, SDIR1 and XERICO were analyzed by Y2H system. For the strong positive interaction control, Krev1 (Rap1A, a member of the Ras family of GTP binding proteins) and RalGDS-wt (the Ral guanine nucleotide dissociator stimulator protein) was used. RalGDS-m1 has weak interaction and RalGDS-m2 has no interaction with Krev1. Yeast MaV203 strains containing the indicated plasmid combinations were grown in SD medium without Leu and Trp to an OD600 of 1.0, and 10 µl aliquots of different dilutions (1, 10⁻¹, 10⁻², 10⁻³) were spotted onto selective and non-selective plates (non-selective medium, SD/−Leu/−Trp; selective medium, SD/−Ura/−Leu/−Trp). The combination of plasmids is indicated on the left and dilution series are indicated at the top. β-galactosidase activity was determined in the MaV203 yeast cells cotransformed with the BD/AD plasmids. Data represent means ± SD from three independent experiments. All the experiments were carried out at least in three replications. (B) Purified GST-AtVBP1 was incubated at 30°C for 2 h with VrUBC1 (E2), yeast E1, Ub, and ATP. Samples were separated by SDS-PAGE, and ubiquitinated proteins were detected by immunoblot analysis using anti-GST antibodies.</p

Yeast complementation and thioester formation of VrUBC1.

<p>(A) For the <i>ubc4/5</i> complementation test, the yeast <i>ubc4/5</i> double mutant was transformed with pYES-GFP or pYES-VrUBC1, and selected on uracil-lacking SD medium plates (SD/−Ura). The wild-type (WT) and the <i>ubc4/5</i> transformants harboring pYES-GFP or pYES-VrUBC1 were grown to an OD600 of 1.0, and 10 µl aliquots of different dilutions (10⁻¹, 10⁻², 10⁻³) were spotted onto SD/−Ura plates and grown for 3 d at 30°C. (B) Thioester formation of VrUBC1. VrUBC1 forms DTT-sensitive ubiquitin adducts. <i>In vitro</i> ubiquitination reactions after 5-min at 37°C were treated with DTT or 4 M urea (-DTT). Reactions were resolved by SDS-PAGE and western blots were performed with with anti-His₆ antibodies.</p

Real-time qRT-PCR analysis of drought-stress maker genes.

<p>Total RNA was extracted from the treated tissues and analyzed by real-time qRT-PCR. Light-grown, 4-week-old plants were dehydrated in a vinyl bag for 6 h. Induction patterns of various ABA- and drought-responsive genes (<i>ABF2</i>, <i>ABF3</i>, <i>ABF4</i>, <i>ABI5</i>, <i>ADH1</i>, and <i>KIN2</i>) were analyzed by real-time qRT-PCR. Data represent the fold induction of each gene by dehydration (6 h) relative to the control treatment (0 h). Mean values from three independent technical replicates were normalized to the levels of an internal control, <i>actin</i> mRNA. Asterisks indicate the significance of the difference from the values between the wild-type and the <i>35S:VrUBC1</i> Arabidopsis transgenic plants as determined by Student’s <i>t</i> test (*0.01 ≤ P<0.05, **P<0.01).</p

Subcellular localization of GFP-fusion proteins and BiFC visualization of the interaction between VrUBC1 and AtVBP1 in <i>Agrobacterium</i>-infiltrated tobacco (<i>Nicotiana benthamiana</i>).

<p>(A) The <i>35S:GFP</i>, <i>35S:GFP-AtVBP1</i>, and <i>35S:AtVrUBC1-GFP</i> constructs were transformed into tobacco leaves via <i>Agrobacterium</i>-infiltration. Protoplasts were isolated from the infiltrated leaves after 36 h. Localization of fusion proteins was visualized by confocal microscopy. (B) YFP^N fusions of VrUBC1 and YFP^C fusion of AtVBP1 were coexpressed in tobacco leaves as previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066056#pone.0066056-Walter1" target="_blank">[66]</a>. Epifluorescence from the interaction between YFP^N fusions of VrUBC1 and YFP^C fusion of AtVBP1 was observed in the nucleus of the protoplasts (scale bar = 20 µm). BiFC of Arabidopsis bZIP63 dimerization is shown as an interaction control as previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066056#pone.0066056-Walter1" target="_blank">[66]</a>. Coexpression of <i>YFP^N-VrUBC1</i>/<i>YFP^C-bZIP63</i> is shown as a negative control.</p

Overexpression of <i>VrUBC1</i>, a Mung Bean E2 Ubiquitin-Conjugating Enzyme, Enhances Osmotic Stress Tolerance in <i>Arabidopsis</i>

<div><p>The ubiquitin conjugating enzyme E2 (UBC E2) mediates selective ubiquitination, acting with E1 and E3 enzymes to designate specific proteins for subsequent degradation. In the present study, we characterized the function of the mung bean <i>VrUBC1</i> gene (<i><u>V</u>igna <u>r</u>adiata <u>UBC</u> 1</i>). RNA gel-blot analysis showed that <i>VrUBC1</i> mRNA expression was induced by either dehydration, high salinity or by the exogenous abscisic acid (ABA), but not by low temperature or wounding. Biochemical studies of VrUBC1 recombinant protein and complementation of yeast <i>ubc4/5</i> by <i>VrUBC1</i> revealed that <i>VrUBC1</i> encodes a functional UBC E2. To understand the function of this gene in development and plant responses to osmotic stresses, we overexpressed <i>VrUBC1</i> in Arabidopsis (<i>Arabidopsis thaliana</i>). The <i>VrUBC1</i>-overexpressing plants displayed highly sensitive responses to ABA and osmotic stress during germination, enhanced ABA- or salt-induced stomatal closing, and increased drought stress tolerance. The expression levels of a number of key ABA signaling genes were increased in <i>VrUBC1</i>-overexpressing plants compared to the wild-type plants. Yeast two-hybrid and bimolecular fluorescence complementation demonstrated that VrUBC1 interacts with AtVBP1 (<i><u>A</u>. <u>t</u>haliana</i><u>V</u>rUBC1 <u>B</u>inding <u>P</u>artner <u>1</u>), a C3HC4-type RING E3 ligase. Overall, these results demonstrate that <i>VrUBC1</i> plays a positive role in osmotic stress tolerance through transcriptional regulation of ABA-related genes and possibly through interaction with a novel RING E3 ligase.</p></div

Northern blotting of <i>VrUBC1</i> in mung bean.

<p><i>VrUBC1</i> RNA expression in mung bean leaves exposed to low temperature (4°C), dehydration, wounding, ABA (100 µM), or salt stress (100 mM NaCl). Twenty micrograms of total RNA was loaded in each lane<b>.</b> Following electrophoresis, RNA was transferred to a nylon membrane and hybridized with a probe specific for <i>VrUBC1</i>. Equal loading of the total RNA (20 µg) was confirmed by EtBr staining, shown as rRNA below the signal panel.</p