Porcine epidemic diarrhoea virus (PEDV) infection activates AMPK and JNK through TAK1 to induce autophagy and enhance virus replication

Autophagy plays an important role in defending against invading microbes. However, numerous viruses can subvert autophagy to benefit their replication. Porcine epidemic diarrhoea virus (PEDV) is an aetiological agent that causes severe porcine epidemic diarrhoea. How PEDV infection regulates autophagy and its role in PEDV replication are inadequately understood. Herein, we report that PEDV induced complete autophagy in Vero and IPEC-DQ cells, as evidenced by increased LC3 lipidation, p62 degradation, and the formation of autolysosomes. The lysosomal protease inhibitors chloroquine (CQ) or bafilomycin A and Beclin-1 or ATG5 knockdown blocked autophagic flux and inhibited PEDV replication. PEDV infection activated AMP-activated protein kinase (AMPK) and c-Jun terminal kinase (JNK) by activating TGF-beta-activated kinase 1 (TAK1). Compound C (CC), an AMPK inhibitor, and SP600125, a JNK inhibitor, inhibited PEDV-induced autophagy and virus replication. AMPK activation led to increased ULK1S777 phosphorylation and activation. Inhibition of ULK1 activity by SBI-0206965 (SBI) and TAK1 activity by 5Z-7-Oxozeaenol (5Z) or by TAK1 siRNA led to the suppression of autophagy and virus replication. Our study provides mechanistic insights into PEDV-induced autophagy and how PEDV infection leads to JNK and AMPK activation.</p

Le Peuple : organe quotidien du syndicalisme

25 juin 19381938/06/25 (A20,N6363)-1938/06/25

Marie-Claire / dir. Jean Prouvost

29 juillet 19381938/07/29 (A0,N74)-1938/07/29.Appartient à l’ensemble documentaire : UnivJeun

Additional file 8: of Development of the body image self-rating questionnaire for breast cancer (BISQ-BC) for Chinese mainland patients

Body Image Self-rating Questionnaire for Breast Cancer (BISQ-BC). (DOC 79 kb

Man with Two Mules

Man in field with two Mules. Writing on the photo or group of photos: 'Mr. Dodson of Tennessee, father of Dr. Dobson who graduated September 17, 1943' Physical description: Black and white print (photograph) 7x11mm.Student by the name George Dobson or "Big D" is listed as a senior in the 1944 Longhorn yearbook, with the home town of College Grove, Tennessee. It is believed that some of the images in this collection are photographs shared by the student's families and are that of animals owned by families farms or ranches

Oxygen and glucose deprivation (OGD)-induced cell injury is attenuated by <i>Spatholobus suberctus</i> Dunn extract (SSCE).

<p><b>A-B.</b> Cell viability by Cell Counting Kit-8 assay. <b>C.</b> Cell apoptosis by flow cytometry. <b>D-E.</b> Protein expression of apoptosis-associated proteins by Western blot analysis. Data are presented as the mean ± SEM. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001. Bcl-2, B cell lymphoma-2; Bax, Bcl-2-associated X protein; P-, pro; C-, cleaved; C/P-, cleaved/pro.</p

Extract of <i>Spatholobus suberctus</i> Dunn ameliorates ischemia-induced injury by targeting miR-494

<div><p>Cerebral stroke is a leading cause of death and permanent disability. The current therapeutic outcome of ischemic stroke (>85% of all strokes) is very poor, thus novel therapeutic drug is urgently needed. <i>In vitro</i> cell model of ischemia was established by oxygen-glucose deprivation (OGD) and in <i>vivo</i> animal model of ischemia was established by middle cerebral artery occlusion (MCAO). The effects of <i>Spatholobus suberctus</i> Dunn extract (SSCE) on OGD-induced cell injury, MCAO-induced neural injury and miR-494 level were all evaluated. The possible target genes were virtually screened utilizing bioinformatics and verified by luciferase assay. Subsequently, the effects of abnormally expressed miR-494 on OGD-induced cell injury and target gene expression were determined. Additionally, whether SSCE affected target gene expression through modulation of miR-494 was studied. Finally, the effects of aberrantly expressed Sox8 on OGD-induced injury and signaling pathways were estimated. SSCE reduced OGD-induced cell injury and ameliorated MCAO-induced neuronal injury, along with down-regulation of miR-494. Then, OGD-induced cell injury was increased by miR-494 overexpression but decreased by miR-494 silence. Sox8 was a target gene of miR-494, and SSCE could up-regulate Sox8 expression via down-regulating miR-494. Afterwards, OGD-induced cell injury was proved to be increased by Sox8 inhibition but reduced by Sox8 overexpression. Finally, OGD-induced inhibition of PI3K/AKT/mTOR and MAPK pathways was further inhibited by Sox8 silence but activated by Sox8 overexpression. SSCE ameliorates ischemia-induced injury both <i>in vitro</i> and <i>in vivo</i> by miR-494-mediated modulation of Sox8, involving activations of PI3K/AKT/mTOR and MAPK pathways.</p></div

The sequence of AaCASPS16.

<p>Predicted amino acid sequence of AaCASPS16 was shown in alignment with Strica homologs from <i>Aedes aegypti</i> (AeCASPS15, AeCASPS16, AeCASPS17 and AeCASPS21) and the caspases from <i>Drosophila melanogaster</i> (Dronc, Dredd, Drice, Decay, Dcp-1, Strica and Damm). The amino acid residues identical among 12 caspases are indicated by white letters within black boxes, the amino acid residues identical among 9 caspases are indicated by black letters within dark gray boxes, the amino acid residues identical among 6 caspases are indicated by black letters within medium gray boxes, and the amino acid residues identical among 3 caspases are indicated by black letters within light gray boxes. The alignment was performed using DNAMAN 7.0. Secondary structures were predicted using JPred3. Underline: catalytic center, black arrow: predicted cleavage site.</p

Sox8 is a target gene of microRNA (miR)-494.

<p><b>A-B.</b> mRNA level of Sox8 by quantitative reverse transcription PCR (qRT-PCR). <b>C-D.</b> Protein level of Sox8 by Western blot analysis. <b>E.</b> Relative luciferase activity by luciferase assay. Data are presented as the mean ± SEM. *, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001. Sox8-WT, pmirGLO vector containing fragments of wild-type rat Sox8 3’UTR; Sox8-Mut, mutant Sox8-WT.</p

Transient expression of AaCASPS16 induced apoptosis in C6/36 cells.

<p>Plasmids expressing C-terminally Flag-tagged AaCASPS16-WT, AaCASPS16-C300A and GFP were transfected into C6/36 cells separately. Caspase inhibitor z-VAD-FMK was added at 2 h before transfection of pIE-AaCASPS16, and proteasome inhibitor MG132 was added at 8 h before cells were harvested. Mock treated cells, GFP expressed C6/36 cells, and AaCASPS16-C300A expressed C6/36 cells were used as controls. At 24 h post transfection, cells were subjected to the following analyses: <b>(A)</b> Photographs of cells were taken under microscope (Scale bar indicated 50 μm). <b>(B)</b> Cell lysates were prepared and were incubated with Ac-DEVD-AFC and subjected to caspase activity assay. Caspase activity was indicated as the changes in relative fluorescence units (RFU) per minute. <b>(C)</b> Cell lysates were subjected to immunoblotting analysis using antibody against Flag and β-tubulin. The data in (B) were presented with the SD from three independent experiments, and statistical significance was calculated by <i>t</i> test, ** <i>P</i> < 0.01. NS: not significant.</p