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Development of temperature and weight after direct oral immunization.
No full text(A) Goats and (B) foxes were directly orally vaccinated with either parental rNDV (n = 3) or RABV G expressing rNDV_GRABV (n = 6). Rectal temperature of goats and foxes as well as weight of foxes was monitored at indicated timepoints after oral vaccination. (DOCX)</p
MMAE changed STING trafficking routes and promoted cGAMP-mediated STING activity.
No full text(A-C and F) HeLa cells stably expressing human STING-GFP were stimulated with cGAMP (8 μM) and/or MMAE (1 μM), or VcMMAE (1 μM) for 2 h. (A and F) Live cells were stained by ER-Tracker Blue-White DPX and LysoTracker Deep Red. (B and C) Cells were fixed, permeabilized, and stained for GM130 (a Golgi protein, red) or tubulin (red). Nuclei were stained with DAPI (blue). All structured illumination microscope (3D-SIM) images are z-stack images. Scale bars, 10 μm. 3D-SIM images was acquired and processed using the Highly Intelligent and Sensitive SIM (HIS-SIM), and Wiener deconvolution was used in reconstructed images. Dashed white boxes in each main image indicate enlarged areas of interest shown below. Co-localization was quantified using Pearson’s correlation coefficient (r), shown on the right of each row of images (n = 50). (D and E) BJ-5ta cells were stimulated with cGAMP (8 μM) with or without MMAE (pre-treatment for 30 min), bafilomycin A1 (BafA1, 100 nM), or brefeldin A (BFA, 1 μM) for 2 h. Total STING protein was quantified by image J software (n = 3 biological replicates). (G and H) STING stability was analyzed by immunoblotting in the absence or presence of cycloheximide (CHX, 50 μg/ml). BJ-5ta cells were treated and analyzed as in (D and E).</p
MMAE specifically enhanced the cGAMP-STING-mediated NF-kB immune response.
No full text(A) A diagram of MMAE promoting STING-mediated NF-kB signaling. (B-E) THP1-Lucia NF-kB (WT, p50 KO and p65 KO) and THP1-Lucia ISG (WT and p65 KO) were treated with cGAMP (0.5 μM) and/or indicated doses of MMAE for 24 h. The fold change of luminescence was normalized to DMSO-treated cells. (F, I and J) HEK293T cells were transiently transfected with STING plasmids (WT, Flag-hSTING (S366A)) for 24 hours. Cells were stimulated with cGAMP (0.5 μM) and/or MMAE (0.1 μM) for 6 h, and cell lysates were analyzed by immunoblotting for the indicated proteins (F and I). Quantification of LC3II/tubulin ratio from three independent experiments (J). (G) THP1-Lucai ISG (STING KO) cells stably expressing (hSTING WT, Flag-hSTING (S366A)) were stimulated with cGAMP and/or MMAE (1 μM) for 12 h. IFNβ production was measured by ELISA analysis. (H) STING was analyzed by immunoblotting in THP1-Lucai ISG (STING KO) cells.</p
Multiple microtubule destabilizers altered STING trafficking pattern during cGAMP-mediated STING activation.
No full text(A) Fluorescent micrograph shows hSTING-GFP vesicle trafficking in HeLa cells. Time-lapse live cell microscopy recording was started 0 min after cGAMP (8 μM) or co-stimulated with MMAE (1 μM) or VcMMAE (1 μM). Selected frames from the movie are shown in A. Scale bars, 5 μm. (B) HeLa cells (hSTING-GFP) were stimulated with cGAMP (8 μM) with or without various microtubule destabilizers and a microtubule stabilizer (paclitaxel, 1 μM) after 2 h, fixed, permeabilized, and stained for tubulin (red). Nuclei were stained with DAPI (blue). Scale bars, 10 μm. (C) Immunoblotting analysis of STING degradation in THP1 cells treated with cGAMP (8 μM) with or without MMAE (1 μM) in the absence or presence of cycloheximide (CHX, 50 μg/ml) for indicated times. Total STING protein was quantified by image J software (n = 3 biological replicates). (TIF)</p
MMAE directly enhanced the cGAMP-STING signaling pathway.
No full text(A) A model showing whether the potentiation effect of MMAE is dependent on the direct STING-IRF3 signal axis or the indirect IFNα/β and its receptors (IFNAR) pathway. (B-I) ISRE reporter activities and STING phosphorylation cascades were measured in response to cGAMP, RO8191 (0.25 μM, an IFNAR2 agonist) or combined with indicated MMAE for 24 h or 6 h in THP1-Lucia ISG cells (WT, STAT1 KO, STAT2 KO and STAT3 KO). The fold changes in luminescent signals were normalized to DMSO-treated cells. The activation of STING signaling was assessed by immunoblotting. Data are representative of three independent experiments. Bars are the mean ± SEM of indicated (n) independent experiments. Significance was determined by one-way ANOVA; *p p p p (TIF)</p
Focus group guide.
No full textBackgroundPeer support in mental health is a low-threshold intervention with increasing evidence for enhancing personal recovery and empowerment of persons living with severe mental health conditions. As peer support spreads globally, there is a growing need for peer support training programmes that work well in different contexts and cultures. This study evaluates the applicability and transferability of implementing a manualised multi-national training programme for mental health peer support workers called UPSIDES from the perspective of different local stakeholders in high-, middle-, and low-income countries.MethodData from seven focus groups across six study sites in Africa (Tanzania, Uganda), Asia (India, Israel), and Europe (Germany 2 sites) with 44 participants (3 service users, 7 peer support workers, 25 mental health staff members, 6 clinical directors and 3 local community stakeholders) were thematically analysed.Results397 codes were identified, which were thematically analysed. Five implementation enablers were identified: (i) Enhancing applicability through better guidance and clarity of training programme management, (ii) provision of sufficient time for training, (iii) addressing negative attitudes towards peer support workers by additional training of organisations and staff, (iv) inclusion of core components in the training manual such as communication skills, and (v) addressing cultural differences of society, mental health services and discrimination of mental health conditions.DiscussionParticipants in all focus groups discussed the implementation of the training and peer support intervention to a greater extent than the content of the training. This is in line with growing literature of difficulties in the implementation of peer support including difficulties in hiring peer support workers, lack of funding, and lack of role clarity. The results of this qualitative study with stakeholders from different mental health settings worldwide emphasises the need to further investigate the successful implementation of peer support training. All results have been incorporated into the manualisation of the UPSIDES peer support training.</div
Untreated patient’s age, cardiac form, and parasitemia levels according to HIV status.
No full textUntreated patient’s age, cardiac form, and parasitemia levels according to HIV status.</p
Rer1 supports Myc-induced growth.
No full text(A-B) Representative images of the wing imaginal discs with hs-FLP induced (at 48 hrs AEL) Actin-FRT-Stop-FRT-Gal4 (AFG)-clones overexpressing GFP and Myc (72 hrs AHS) in either wild-type (A) or rer1+/–background (B). (C) Quantification of p-eIF2α shows higher levels in AFG:: Myc, GFP; rer1+/–(N = 15 wing discs) as compared to AFG:: Myc, GFP in WT background (N = 15 wing discs). (D) Quantification of GFP positive area shows reduction in AFG:: Myc, GFP; rer1+/–(N = 21 wing discs) as compared to AFG:: Myc, GFP in WT background (N = 18 wing discs). Statistical analyses in C and D were performed using the Two-tailed Welch’s unpaired t-test (**** p(E-G) Schematic representation of the involvement of Rer1 in the regulation of proteostasis and its role in supporting Myc-induced overgrowth. (E) Rer1 plays a homeostatic role in the regulation of protein quality providing cytoprotection in the wild-type cells. (F) Cells lacking Rer1 have proteotoxic stress due to improper cytoprotection. This leads to high cytotoxicity downstream of UPR activation, ultimately leading to the competitive elimination of cells identified as losers. (G) Myc-overexpression increases gene expression leading to high proteotoxic stress. Rer1 levels are upregulated upon Myc-overexpression to maintain higher demand of proteostasis, thereby supporting overgrowth. Also see S11 Fig.</p
Viral shedding levels in H7N7-LP<sub>FLAGtag</sub>/H7N7-HP<sub>HAtag</sub> co-inoculated chickens and ducks as determined by endpoint titration assay in MDCK cells.
No full textInfectious virus quantification in oropharyngeal (OP; solid lines) and cloacal (CL; dotted lines) swabs from H7N7-LPFLAGtag/H7N7-HPHAtag co-inoculated chickens (A) and ducks (B). Infectious titers were determined by endpoint titration in MDCK cells and expressed as log10 TCID50/mL (viral RNA levels and plaque forming units are depicted in Figs 4 and 5). (TIF)</p
Fig 7 -
No full textChromatographic profiles of VOCs of (A) strain IALR1619 incubated for 72 h in LB medium and (B) uninoculated LB medium. The peak corresponds to 1-undecene.</p