Increased Expression of Interleukin-6 Family Members and Receptors in Urinary Bladder with Cyclophosphamide-Induced Bladder Inflammation in Female Rats

Recent studies suggest that janus-activated kinases–signal transducer and activator of transcription signaling pathways contribute to increased voiding frequency and referred pain of cyclophosphamide (CYP)-induced cystitis in rats. Potential upstream chemical mediator(s) that may be activated by CYP-induced cystitis to stimulate JAK/STAT signaling are not known in detail. In these studies, members of the interleukin (IL)-6 family of cytokines including, leukemia inhibitory factor (LIF), IL-6, and ciliary neurotrophic factor (CNTF) and associated receptors, IL-6 receptor (R) α, LIFR, and gp130 were examined in the urinary bladder in control and CYP-treated rats. Cytokine and receptor transcript and protein expression and distribution were determined in urinary bladder after CYP-induced cystitis using quantitative, real-time polymerase chain reaction (Q-PCR), western blotting, and immunohistochemistry. Acute (4 h; 150 mg/kg; i.p.), intermediate (48 h; 150 mg/kg; i.p.), or chronic (75 mg/kg; i.p., once every 3 days for 10 days) cystitis was induced in adult, female Wistar rats with CYP treatment. Q-PCR analyses revealed significant (p ≤ 0.01) CYP duration- and tissue- (e.g., urothelium, detrusor) dependent increases in LIF, IL-6, IL-6Rα, LIFR, and gp130 mRNA expression. Western blotting demonstrated significant (p ≤ 0.01) increases in IL-6, LIF, and gp130 protein expression in whole urinary bladder with CYP treatment. CYP-induced cystitis significantly (p ≤ 0.01) increased LIF-immunoreactivity (IR) in urothelium, detrusor, and suburothelial plexus whereas increased gp130-IR was only observed in urothelium and detrusor. These studies suggest that IL-6 and LIF may be potential upstream chemical mediators that activate JAK/STAT signaling in urinary bladder pathways

The significance of lifeworld and the case of hospice

Questions on what it means to live and die well are raised and discussed in the hospice movement. A phenomenological lifeworld perspective may help professionals to be aware of meaningful and important dimensions in the lives of persons close to death. Lifeworld is not an abstract philosophical term, but rather the opposite. Lifeworld is about everyday, common life in all its aspects. In the writings of Cicely Saunders, known as the founder of the modern hospice movement, facets of lifeworld are presented as important elements in caring for dying patients. Palliative care and palliative medicine today are, in many ways, replacing hospices. This represents not only a change in name, but also in the main focus. Hospice care was originally very much about providing support and comfort for, and interactions with the patients. Improved medical knowledge today means improved symptomatic palliation, but also time and resources spent in other ways than before. Observations from a Nordic hospice ward indicate that seriously ill and dying persons spend much time on their own. Different aspects of lifeworld and intersubjectivity in the dying persons’ room is presented and discussed

ϒ production in p–Pb collisions at √sNN=8.16 TeV

ϒ production in p–Pb interactions is studied at the centre-of-mass energy per nucleon–nucleon collision √sNN = 8.16 TeV with the ALICE detector at the CERN LHC. The measurement is performed reconstructing bottomonium resonances via their dimuon decay channel, in the centre-of-mass rapidity intervals 2.03 < ycms < 3.53 and −4.46 < ycms < −2.96, down to zero transverse momentum. In this work, results on the ϒ(1S) production cross section as a function of rapidity and transverse momentum are presented. The corresponding nuclear modification factor shows a suppression of the ϒ(1S) yields with respect to pp collisions, both at forward and backward rapidity. This suppression is stronger in the low transverse momentum region and shows no significant dependence on the centrality of the interactions. Furthermore, the ϒ(2S) nuclear modification factor is evaluated, suggesting a suppression similar to that of the ϒ(1S). A first measurement of the ϒ(3S) has also been performed. Finally, results are compared with previous ALICE measurements in p–Pb collisions at √sNN = 5.02 TeV and with theoretical calculations.publishedVersio

Centrality evolution of the charged-particle pseudorapidity density over a broad pseudorapidity range in Pb-Pb collisions at root s(NN)=2.76TeV

Peer reviewe

Upregulation of vascular endothelial growth factor isoform VEGF-164 and receptors (VEGFR-2, Npn-1, and Npn-2) in rats with cyclophosphamide-induced cystitis

Regulation of the VEGF-VEGF receptor system was examined in the urinary bladder after acute (2–48 h) and chronic (10 days) cyclophosphamide (CYP)-induced cystitis. ELISAs demonstrated significant (P ≤ 0.01) upregulation of VEGF in whole urinary bladder with acute and chronic CYP-induced cystitis; however, the magnitude of increase was greater after acute (2–4 h) cystitis. Immunohistochemistry for VEGF immunoreactivity revealed a significant (P ≤ 0.05) increase in VEGF immunoreactivity in the urothelium, suburothelial vasculature, and detrusor smooth muscle with acute (4 and 48 h) CYP treatment. RT-PCR identified the isoform VEGF-164, the VEGF receptor VEGFR-2, and the VEGF co-receptors neuropilin (Npn)-1 and Npn-2 in the urinary bladder. Quantitative PCR demonstrated upregulation of VEGF-164 transcript with acute and chronic CYP-induced cystitis, but VEGFR-2, Npn-1, and Npn-2 transcripts were upregulated (P ≤ 0.01) in whole bladder only with chronic CYP-induced cystitis. Additional studies demonstrated regulation of VEGF transcript expression in the urinary bladder by nerve growth factor (NGF) in a novel line of NGF-overexpressing mice. These studies demonstrated that urinary bladder inflammation and NGF regulate the VEGF-VEGF receptor system in the urinary bladder. Functional role(s) for the VEGF-VEGF receptor system in urinary bladder inflammation remain to be determined

Global Analysis of the Sporulation Pathway of <i>Clostridium difficile</i>

<div><p>The Gram-positive, spore-forming pathogen <i>Clostridium difficile</i> is the leading definable cause of healthcare-associated diarrhea worldwide. <i>C. difficile</i> infections are difficult to treat because of their frequent recurrence, which can cause life-threatening complications such as pseudomembranous colitis. The spores of <i>C. difficile</i> are responsible for these high rates of recurrence, since they are the major transmissive form of the organism and resistant to antibiotics and many disinfectants. Despite the importance of spores to the pathogenesis of <i>C. difficile</i>, little is known about their composition or formation. Based on studies in <i>Bacillus subtilis</i> and other <i>Clostridium</i> spp., the sigma factors σ^F, σ^E, σ^G, and σ^K are predicted to control the transcription of genes required for sporulation, although their specific functions vary depending on the organism. In order to determine the roles of σ^F, σ^E, σ^G, and σ^K in regulating <i>C. difficile</i> sporulation, we generated loss-of-function mutations in genes encoding these sporulation sigma factors and performed RNA-Sequencing to identify specific sigma factor-dependent genes. This analysis identified 224 genes whose expression was collectively activated by sporulation sigma factors: 183 were σ^F-dependent, 169 were σ^E-dependent, 34 were σ^G-dependent, and 31 were σ^K-dependent. In contrast with <i>B. subtilis</i>, <i>C. difficile</i> σ^E was dispensable for σ^G activation, σ^G was dispensable for σ^K activation, and σ^F was required for post-translationally activating σ^G. Collectively, these results provide the first genome-wide transcriptional analysis of genes induced by specific sporulation sigma factors in the Clostridia and highlight that diverse mechanisms regulate sporulation sigma factor activity in the Firmicutes.</p></div

<i>C. difficile sigF</i>⁻, <i>sigE⁻</i>, <i>sigG⁻</i>, and <i>sigK⁻</i> mutants are arrested at different stages of spore formation.

<p>Transmission electron microscopy (TEM) of wildtype, <i>spo0A⁻</i>, <i>sigF</i><b><i>⁻</i></b>, <i>sigE</i><b><i>⁻</i></b>, <i>sigG</i><b><i>⁻</i></b>, and <i>sigK</i><b><i>⁻</i></b> strains at 18 hrs of growth on sporulation media. The forespore regions of wild type (WT), <i>sigG⁻</i>, and <i>sigK⁻</i> strains, and an electron-translucent region within the <i>sigF⁻</i> mutant mother cell cytosol, are shown on the right. Black triangles indicate regions that resemble coat layers, while white triangles indicate regions consistent with cortex. Scale bars represent 500 nm. Inset scale bars represent 250 nm.</p

Comparison of Spo0A-dependent gene expression in wildtype and sporulation sigma factor mutants.

<p>Heat map representation of the genes that were downregulated by ≥4-fold with an adjusted p-value of ≤10⁻⁵ in the <i>spo0A</i>⁻ strain relative to wild type. Expression levels of these genes in wildtype (WT), <i>sigF⁻</i>, <i>sigE⁻</i>, <i>sigG⁻</i>, and <i>sigK⁻</i> strains relative to <i>spo0A⁻</i> were centered, scaled, and mapped to a red-green color scale, with green indicating that the gene was upregulated in the strain relative to the other strains, and red indicating that the gene was downregulated relative to the centered expression level. σ^E-regulated genes (blue), σ^K-regulated genes (purple), σ^G-regulated genes (orange), σ^F-dependent genes (green) are colored as indicated, and clusters of coordinately regulated genes are bracketed. Genes colored in black were identified as depending only on Spo0A for expression.</p

Western blot analyses of proteins encoded by genes induced by specific sigma factors during sporulation.

<p>Western blot analyses of proteins encoded by genes identified as being upregulated during sporulation by specific sigma factors. Wildtype (WT), <i>spo0A⁻</i>, <i>sigF⁻</i>, <i>sigE⁻</i>, <i>sigG⁻</i>, and <i>sigK⁻</i> strains were grown on sporulation media for 18 hrs. SleC undergoes multiple processing steps <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003660#pgen.1003660-Adams1" target="_blank">[76]</a>, <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003660#pgen.1003660-Okamura1" target="_blank">[85]</a>, but only the pro-SleC form is shown. Spo0A was used as a loading control.</p