Journal: The Journal of General Virology

Article Title: Increased serum sialic acid is associated with morbidity and mortality in a murine model of dengue disease

doi: 10.1099/jgv.0.001319

Figure Lengend Snippet: Sialic acid and NS1 levels in serum of DENV-infected mice increase proportionally with infectious dose and are correlated with DENV-induced morbidity. Ifnar −/− mice were infected i.v. with increasing doses of DENV2 D220, and blood samples were collected on day 3 post-infection for serum testing by ELISA. Quantification of (a) Sia and (b) NS1 levels in serum from DENV-infected mice (mean±sd). Correlation analysis between DENV-induced (c) morbidity and Sia, (d) morbidity and NS1, and (e) Sia and NS1 levels in serum. Signs of morbidity were scored using a standardized scale, where 1=healthy; 2=mild signs of lethargy, some fur ruffling, no hunching; 3=fur ruffling, hunched, mild signs of lethargy; 4=ruffled, hunched and lethargic, limited morbidity; 5=moribund or dead. Data were combined from two experiments. ADE, antibody-dependent enhancement; p.f.u., plaque-forming units.

Article Snippet: Total Sia levels (conjugated and free) in mouse sera were measured using a Mouse Sialic Acid ELISA kit (MyBioSource) as per the manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: The Journal of General Virology

Article Title: Increased serum sialic acid is associated with morbidity and mortality in a murine model of dengue disease

doi: 10.1099/jgv.0.001319

Figure Lengend Snippet: Circulating levels of sialic acid and NS1 are associated with DENV infection outcome. Ifnar −/− mice were infected i.v. with different doses of DENV2 D220, and survival was monitored for 10 days. These are the same mice as those tested in Fig. 1. (a) A Kaplan–Meier survival curve is shown. (b) Sia and (c) NS1 levels in serum were evaluated by ELISA on day 3 post-infection and plotted in relation to DENV infection outcome (mean±sd), with significantly higher amounts being found in animals that succumbed to the infection, with P-values as indicated (two-sided Student t-test). Data were combined from two similar experiments. ADE, antibody-dependent enhancement; p.f.u., plaque-forming units.

Article Snippet: Total Sia levels (conjugated and free) in mouse sera were measured using a Mouse Sialic Acid ELISA kit (MyBioSource) as per the manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: The Journal of General Virology

Article Title: Increased serum sialic acid is associated with morbidity and mortality in a murine model of dengue disease

doi: 10.1099/jgv.0.001319

Figure Lengend Snippet: DENV-induced morbidity and mortality in a murine model of dengue disease are correlated with increasing levels of Sia and NS1 in circulation. (a) Ifnar —/— mice were infected with DENV2-D220, and 3 days post-infection, circulating levels of Sia were measured by (b) ELISA and (c) a newly developed LC-MRM-MS assay. The latter approach showed that serum levels of free N-glycolylneuraminic acid (Neu5Gc), a form of Sia found in most non-human mammals, were significantly higher in DENV-infected mice. (d) High levels of NS1 and Sia were associated with increased mortality in DENV-infected mice. These findings indicate that DENV infection leads to disruption of EGL components, such as Sia, which can be measured in serum and could used as potential biomarkers for severe dengue disease in humans.

Article Snippet: Total Sia levels (conjugated and free) in mouse sera were measured using a Mouse Sialic Acid ELISA kit (MyBioSource) as per the manufacturer’s instructions.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Disruption

Journal: Shock (Augusta, Ga.)

Article Title: The Protective Effect of a Short Peptide Derived from Cold-inducible RNA-binding Protein in Renal Ischemia-Reperfusion Injury

doi: 10.1097/SHK.0000000000000988

Figure Lengend Snippet: Serum was collected 24 h after RIR and used to measure (A) TNF-α, (B) IL-6, (C) IL-1β using ELISA. Data are expressed as mean ± SEM (sham: n = 4 mice; RIR + vehicle: n = 5 mice; RIR + vehicle: n = 5 mice) and compared by one-way analysis of variance and Student-Newman-Keuls method. *P < 0.05 versus sham; #P < 0.05 versus vehicle. RIR, renal ischemia-reperfusion; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Serum TNF-α, IL-6, and IL-1β were determined with an ELISA kit, specific to mouse TNF-α, IL-6, and IL-1β (BD Biosciences, San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Low-intensity pulsed ultrasound triggers a beneficial neuromodulation in dementia mice with chronic cerebral hypoperfusion via activation of hippocampal Fndc5/irisin signaling

doi: 10.1186/s12967-022-03824-7

Figure Lengend Snippet: Boosting hippocampal Fndc5/irisin alleviate hippocampal damage and cognitive deficits following experimental VaD. A Experimental outline. Forced expression of Fndc5 in hippocampus of adult C57BL/6 mice using adenovirus brain infection, while enhancement of hippocampal irisin level by bilateral intrahippocampal injection of recombinant irisin on day 22 after BCAS. B Hippocampal Fndc5 mRNA expression and irisin were respectively detected using qPCR and ELISA on day 28 after BCAS. C Hippocampal synaptic plasticity was analyzed using Patch Clamp on day 28 after BCAS. The average traces of fEPSP in hippocampal slices. D The data of fEPSP at 120 min. E – G Cognitive evaluation were performed on day 28 after BCAS. The errors (numbers of entries into incorrect maze arms) in two-days radial arm water maze test were continuously measured in each block ( E ). The errors in last block were counted in a histogram ( F ). Exploratory time on old or new object in novel object recognition test were illustrated in histogram ( G ). H , I Hippocampal level of TGF-β and IL-10, were determined using ELISA on day 28 after BCAS. J The percentage of freezing time obtained on day 28 after BCAS in contextual fear test were illustrated in histogram. Sham, sham-operated mice; BCAS, mice were subjected to BCAS injury; AdFndc5 or AdGFP, adult BCAS mice were infected using intracerebroventricular injection with an adenoviral vector designed to express Fndc5 or GFP; Irisin, adult BCAS mice with bilateral intrahippocampal injection of recombinant irisin. (n = 8 per group. All data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns means no statistical significance)

Article Snippet: The concentration of irisin in serum, brain tissue homogenate or culture supernatant were detected using the irisin ELISA kit obtained from Phoenix pharmaceuticals (Burlingame, USA). and were obtained from Phoenix pharmaceuticals (Burlingame, USA). transforming growth factor (TGF)-β, and interleukin (IL)-10 were measured with mouse TGF-β and IL-10 ELISA Kits (Aviscera Bioscience, Santa Clara, CA, USA).

Techniques: Expressing, Infection, Injection, Recombinant, Enzyme-linked Immunosorbent Assay, Patch Clamp, Blocking Assay, Plasmid Preparation

Journal: Journal of Translational Medicine

Article Title: Low-intensity pulsed ultrasound triggers a beneficial neuromodulation in dementia mice with chronic cerebral hypoperfusion via activation of hippocampal Fndc5/irisin signaling

doi: 10.1186/s12967-022-03824-7

Figure Lengend Snippet: Fndc5/irisin mediates the neurorestorative effects of LIPUS on experimental VaD. A Experimental outline. Knockdown the expression of hippocampal Fndc5 using shRNA before 7days of BCAS. Transcranial LIPUS was applied 24 h after BCAS and subsequently daily with a stimulation time of 5 min at an ultrasound pressure of 0.51 MPa for a period of 28 days. B , C Fndc5 mRNA expression and irisin concentration in hippocampus of adult mice were respectively detected using qPCR and ELISA on day 28 after BCAS. D Hippocampal synaptic plasticity was analyzed using Patch Clamp, average traces of fEPSP in hippocampal slices collected on day 28 after BCAS. E The data of fEPSP at 120 min. F , G Hippocampal TGF-β and IL-10 were determined using ELISA on day 28 after BCAS. H Exploratory time on old or new object in novel object recognition test were illustrated in histogram. I The errors in radial arm water maze test were continuously measured in each block on day 28 after BCAS. J The data of errors in last block were counted in a histogram. K Global Fndc5 knock-out mice (F5KO) was employed to mechanism exploration, contextual fear test was used to assess cognitive deficit on day 28 after BCAS. the percentage of freezing time obtained on day 28 after BCAS were illustrated in histogram. Sham, sham-operated mice; BCAS, mice were subjected to BCAS injury; LIPUS, BCAS adult mice with LIPUS treatment; shFndc5 or shLuc, LIPUS-treated adult BCAS mice with intrahippocampal injection with lentiviral particles expressing shRNA against murine Fndc5 or luciferase. (n = 8 per group. All data are expressed as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns means no statistical significance)

Article Snippet: The concentration of irisin in serum, brain tissue homogenate or culture supernatant were detected using the irisin ELISA kit obtained from Phoenix pharmaceuticals (Burlingame, USA). and were obtained from Phoenix pharmaceuticals (Burlingame, USA). transforming growth factor (TGF)-β, and interleukin (IL)-10 were measured with mouse TGF-β and IL-10 ELISA Kits (Aviscera Bioscience, Santa Clara, CA, USA).

Techniques: Expressing, shRNA, Concentration Assay, Enzyme-linked Immunosorbent Assay, Patch Clamp, Blocking Assay, Knock-Out, Injection, Luciferase