29 research outputs found

    Effects of exogenous adenosine on ileal contractility during HSV-1 infection.

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    <p>(A) Concentration–response curves to adenosine in rat ileum from sham and infected rats at 1 and 6 weeks (w) after HSV-1 intragastric inoculum. Data are means ± SEM and are expressed as gram tension/gram dry tissue weight (four tissue samples from n = 6 animals for each experimental group). (B) Excitatory effects induced by 1 mM adenosine before and after treatment with atropine, a muscarinic receptor blocker (1 µM) or tetrodotoxin, a Na<sup>+</sup> voltage-gated neural channel blocker (TTX, 1 µM). Data are means ± SEM and are expressed as g tension/g dry tissue weight (four tissue samples from n = 6 animals for each experimental group). *P<0.05 compared to the adenosine response in sham rats. <sup>#</sup>P<0.05 compared to the adenosine response in infected rats at 1 week post-intragastric inoculum.</p

    Effects of selective A<sub>1</sub> or A<sub>2A</sub> receptor antagonist on adenosine-induced ileal contraction during HSV-1 infection.

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    <p>Excitatory effects induced by adenosine at concentration of 1 mM before and after treatment with DPCPX (10 nM, panel A) or ZM241385 (10 nM, panel B) in rat ileum from sham and infected rats at 1 and 6 weeks (w) after HSV-1 intragastric administration. Data are means ± SEM and are expressed as g tension/g dry tissue weight (four tissue samples from n = 6 animals for each experimental group). *P<0.05 when compared to the response to adenosine in sham rats. <sup>#</sup>P<0.05 when compared to the response to adenosine in infected rats at 1 week post intragastric inoculum. <sup>§</sup>P<0.05 when compared to the response to adenosine in infected rats at 6 week post intragastric inoculum.</p

    Distribution of the enzymes CD73 and ADA in the neuromuscular layer of rat ileum during HSV-1 infection.

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    <p>Representative immunofluorescence staining of CD73 and ADA in ileal sections from sham and HSV-1-treated rats 1 and 6 weeks (w) after intragastric administration (n = 3 animals for each experimental group). Micrographs showed the presence of CD73 and ADA in the ileal tissue stained with anti-CD73 and anti-ADA antibodies. The boxed areas (panel A) are enlarged at right (panel B) to show the distribution of the CD73 and ADA in the neuromuscular layer. Scale bar: 50 µm (panel A); 25 µm (panel B). CM, circular muscle; LM, longitudinal muscle; *, myenteric ganglia.</p

    PCR primers and probe sequences, PCR annealing temperature, and size of the amplicons.

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    <p>TK, thymidine kinase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LAT, latency associated transcript; ICP4, infected cell protein 4.</p

    Nature of HSV-1 infection in the ENS.

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    <p>RNA was purified from myenteric ganglia of sham and HSV-1 infected rats at 1 and 6 weeks (w) after HSV-1 intragastric administration and the relative expression of HSV-1 LATs (A), and of the very early gene ICP4 (B) was determined by qPCR. Glyceraldehyde-3-phosphate dehydrogenase was used as internal control. Data are expressed as means ± SEM (two tissue samples from n = 3 animals for each experimental group). *P<0.01 compared to sham rats; <sup>#</sup>P<0.01 compared to infected rats at 1 week post-intragastric administration.</p

    Expression of the enzyme ADA in rat ileum during HSV-1 infection.

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    <p>Representative Western blot and densitometric analysis of ADA expression levels in whole thickness ileum without mucosa from sham and HSV-1-treated rats 1 and 6 weeks (w) after intragastric administration (three tissue samples from n = 3 animals for each experimental group). β-actin was used to normalize loading. *P<0.05 compared to sham animals.</p

    Effects of selective adenosine A<sub>1</sub> or A2<sub>A</sub> receptor agonist on ileal contractility during HSV-1 infection.

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    <p>Excitatory effects induced by 10 µM CPA (A) or 10 µM CGS-21680 (B) before and after treatment with atropine (1 µM) or tetrodotoxin (TTX, 1 µM) in rat ileum from sham and infected rats at 1 and 6 weeks (w) after HSV-1 intragastric administration. Data are means ± SEM and are expressed as g tension/g dry tissue weight (four tissue samples from n = 6 animals for each experimental group). *P<0.05 compared to the agonist response in sham rats. <sup>#</sup>P<0.05 compared to the response of the respective agonist in infected rats at 1 week post-intragastric inoculum. <sup>§</sup>P<0.05 compared to the respective agonist in infected rats at 6 weeks post-intragastric inoculum.</p

    Distribution of adenosine receptors A<sub>1</sub>, A<sub>2A</sub>, A<sub>2B</sub> and A<sub>3</sub> receptors in the neuromuscular layer of rat ileum during HSV-1 infection.

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    <p>Representative immunofluorescence staining of adenosine receptors A<sub>1</sub>, A<sub>2A</sub>, A<sub>2B</sub> and A<sub>3</sub> in ileal sections from control (sham) and HSV-1-treated rats 1 and 6 weeks (w) after intragastric administration. Immunofluorescence analyses revealed expression of all four adenosine receptors in smooth muscle layers and myenteric ganglia of control ileum as well as changes in their immunoreactivity, both at the ganglionic level and in smooth muscle layers over the time-course of viral infection. The boxed areas (panel A) are enlarged at right (panel B) to show the distribution of the adenosine receptors in the neuromuscular layer. Scale bar: 50 µm (panel A); 25 µm (panel B). CM, circular muscle; LM, longitudinal muscle; *, myenteric ganglia.</p

    Inflammatory markers in LMMP during HSV-1 infection.

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    <p>IL-2 (A), TNF-α (B), IFN-γ (C) levels were determined in the LMMPs of sham and HSV-1 infected rats at 1 and 6 weeks (w) after HSV-1 intragastric inoculum (two tissue samples from n = 3 animals for each experimental group). Data are expressed as means ± SEM pg/mg protein.*P<0.01 compared to sham rats; <sup>#</sup>P<0.01 compared to infected rats at 1 week post-intragastric administration.</p

    Scheme for the experimental course of the <i>S</i>. <i>boulardii</i> CNCM I-745 supplementation in HSV-1 infected mice.

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    <p>Mice were intranasally (IN) inoculated with Vero cell lysate (vehicle) or HSV-1 (10<sup>2</sup> PFU). After 4 weeks, mice received an intragastric inoculum (IG) of Vero cell lysate (vehicle) or HSV-1 (10<sup>8</sup> PFU). Mice were randomly allocated to receive 100 μL of either phosphate buffered saline (vehicle) or <i>S</i>. <i>boulardii</i> CNCM I-745 by daily oral gavage starting 4 weeks after IG viral inoculum. Animals were sacrificed 10 weeks after IG viral inoculum. n = 8 mice for each experimental group.</p
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