Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists-1

<p><b>Copyright information:</b></p><p>Taken from "Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists"</p><p>BMC Clinical Pathology 2006;6():3-3.</p><p>Published online 15 Feb 2006</p><p>PMCID:PMC1382240.</p><p>Copyright © 2006 Crockett et al; licensee BioMed Central Ltd.</p> mg/kg), prior to the onset of ischemia (90 min), followed by 3, 6, and 24 h of reperfusion (IR). Nicotine or DMPP treatment was administered 15 min prior to the onset of ischemia and repeated after 3 and 6 h of reperfusion. Sham animals received the same pretreatment as the IR group, followed by sham operation (n = 3 mice each group). "Control" group represents mice that received no drug treatment, and no surgical operation of IR/sham protocol was performed (n = 3 mice). Values are expressed as means ± SEM of = 8 to 13 mice per saline or DMPP group, and n = 4–8 per nicotine group.< 0.05, IR group vs. sham-operated group. < 0.05, Saline-treated IR group vs. DMPP-or nicotine-treated IR group

Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists-5

<p><b>Copyright information:</b></p><p>Taken from "Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists"</p><p>BMC Clinical Pathology 2006;6():3-3.</p><p>Published online 15 Feb 2006</p><p>PMCID:PMC1382240.</p><p>Copyright © 2006 Crockett et al; licensee BioMed Central Ltd.</p>h vehicle (i.e. normal saline), DMPP (1 mg/kg), or nicotine (1 mg/kg), prior to the onset of ischemia (90 min), followed by 3 h of reperfusion (IR). The PCR products were run on 2.5% agarose gel, stained with ethidium bromide, and then visualized by ultraviolet illumination. GAPDH was used as a housekeeping gene. Blots shown are representative of four separate experiments with similar results

Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists-0

<p><b>Copyright information:</b></p><p>Taken from "Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists"</p><p>BMC Clinical Pathology 2006;6():3-3.</p><p>Published online 15 Feb 2006</p><p>PMCID:PMC1382240.</p><p>Copyright © 2006 Crockett et al; licensee BioMed Central Ltd.</p>set of ischemia (90 min), followed by 3 h of reperfusion. Values are expressed as means ± SEM of = 4 mice each group. *< 0.05, Saline-treated group vs. DMPP-treated group

Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists-2

<p><b>Copyright information:</b></p><p>Taken from "Protection of early phase hepatic ischemia-reperfusion injury by cholinergic agonists"</p><p>BMC Clinical Pathology 2006;6():3-3.</p><p>Published online 15 Feb 2006</p><p>PMCID:PMC1382240.</p><p>Copyright © 2006 Crockett et al; licensee BioMed Central Ltd.</p>, prior to the onset of ischemia (90 min), followed by various reperfusion times (IR). Sham animals received the same pretreatment as the IR group, followed by sham operation. The ischemic liver sections were prepared and stained with H&E. The top row images represent the sham mice, which show normal hepatic histology. The following rows represent mice that were subjected to I/R. A pattern of reperfusion damage is evident by necrosis of hepatocytes in the pericentral and midzonal regions, with relative sparing of the periportal areas. Note the presence of neutrophils in the midzonal region around the central vein. Images are representative of n = 13 mice per saline or DMPP, and n = 8 per nicotine group

Synthetic Reconstruction of icGD03-S

<div><p>(A) Unrooted phylogenetic gene tree of 35 SARS isolates ranging from early, middle, and late phases of the 2002–2003 epidemic to 2003–2004 animal isolates. Branch confidence values are shown as posterior probabilities. The three human isolates that fall within the cluster otherwise isolated from animals (shown in boxes), GZ0402, GD03, and GZ0401, may represent infections in which a human acquired the virus from a Himalayan palm civet.</p> <p>(B) The GDO3-S glycoprotein. Amino acid changes unique to the GDO3-S with the GDO3-S amino acid listed on the left and the corresponding Urbani to the right. The GDO3-S amino acid changes are shown in relation to the S1 and S2 subunits, the receptor binding domain (RBD), heptad repeats one (HR1) and two (HR2), the transmembrane domain (TM), and known neutralizing epitopes. Two mutations that arose during tissue culture passage of the chimeric icGDO3-S are shown in red.</p> <p>(C) Growth curves of the Urbani strain of SARS-CoV (diamond, solid line), the recombinant Urbani icSARS (squares, dashed line), and the recombinant chimeric virus icGDO3-S (triangles, dotted line) in human airway epithelial cells.</p> <p>(D) Comparing growth of icSARS-CoV to icGDO3-S in the lungs of mice. Six-week-old female BALB/C mice were infected with icSARS-CoV or icGDO3-S (<i>n</i> = 5 per group). The individual titer of each mouse is represented by a filled circle, and the mean titer of the group is represented as a solid bar.</p></div

Pathogenic Findings Following Homologous Challenge

<p>Light photomicrographs of representative histologic lung sections (<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030525#pmed-0030525-t001" target="_blank">Table 1</a>, experiment 2) taken from an untreated control mouse (A), a VRP-N–vaccinated mouse (B) and (D), a VRP-HA–vaccinated mouse (C), a VRP-S–vaccinated mouse (E), and a VRP-S– and VRP-N–treated mouse (F). No histopathology was evident in (A). A marked mixed inflammatory infiltrate composed mainly of mononuclear leukocytes (lymphocytes and plasma cells) and widely scattered eosinophils are evident in the perivascular and peribronchiolar interstitium (asterisk) in (B). Similar inflammatory cells are also present in bronchiolar (br) airways and alveolar airspaces along with enlarged and vacuolated alveolar macrophages (arrows). The box in (B) denotes the site of the light photomicrograph (D) that was taken at a higher magnification to better illustrate the lymphoplasmacytic inflammatory cell infiltrate with lesser numbers of eosinophils (arrows). Similar, but slightly less severe, perivascular inflammatory infiltrates (asterisk) are also present in (F), but without accompanying alveolitis. Minimal lymphoplasmacytic cell accumulations around the pulmonary arteriole (a) are evident in (C) and (E). All tissues were stained with hematoxylin and eosin. Bars denote the scale of the magnification. a, pulmonary arteriole; ap, alveolar parenchyma; br, bronchiolar lumen; e, surface epithelium of the bronchiole.</p

An 80% Plaque Reduction Neutralization Titers (PRNT₈₀) for VRP-S and VRP-S+N Hyperimmune Serum

<div><p>(A) Mice vaccinated young: icSARS-CoV (left) and icGDO3-S (right) PRNT₈₀ for VRP-S immune serum (experiment 2) collected at 5 wk post-boost (<i>n</i> = 5) and 53 wk post-boost (<i>n</i> = 8).</p> <p>(B) Mice vaccinated old: icSARS (left) and icGDO3-S (right) PRNT₈₀ values for VRP-S and VRP-S+N immune serum (experiment 4) at 12 and 29 wk post-boost (<i>n</i> = 6 for icSARS; <i>n</i> = 5 for icGDO3-S). The PRNT₈₀ values for individual animals are show as black circles, and the mean value is shown as a solid bar. The limits of detection (1:1,600 upper and 1:100 lower) are represented by horizontal dotted lines.</p></div

Kinetics of VRP-N–Associated Inflammation

<p>Light photomicrographs of lung sections taken from VRP-HA– and VRP-N–vaccinated mice harvested at days 2, 4, 7, and 14 post–icSARS-CoV challenge (<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030525#pmed-0030525-t001" target="_blank">Table 1</a>, experiment 5). Representative lung sections (200× magnification) comparing pulmonary inflammation between VRP-HA–vaccinated (A), (C), (E), and (G) and VRP-N–vaccinated (B), (D), (F), and (H) mice. Enhanced inflammation was evident by day 2 (A) and (B) in some VRP-N–vaccinated animals relative to lung sections of VRP-HA–inoculated mice. By day 4 post-infection (C) and (D), increased inflammation in VRP-N–vaccinated animals was widely apparent and was maintained through days 7 (E) and (F) and 14 (G) and (H).</p

Identifying Eosinophils among Inflammatory Infiltrates

<p>The 400× magnification comparing eosinophil infiltration within the lung sections of VRP-HA–vaccinated (A), (C), (E), and (G) and VRP-N–vaccinated (B), (D), (F), and (H) mice (<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030525#pmed-0030525-t001" target="_blank">Table 1</a>, Experiment 5). At day 2 post-infection (A) and (B), eosinophils are rarely evident in the lungs of either VRP-HA (A) or VRP-N (B) mice. Day 4 post-infection (C) and (D), extensive eosinophils (yellow arrows) are present within the lungs of VRP-N–vaccinated mice. Widespread eosinophils are seen at day 7 post-challenge in VRP-N–vaccinated (F), but not VRP-HA–vaccinated (E) mice. By day 14 (G) and (H), eosinophils are rarely found among inflammatory cells of VRP-N–vaccinated mice. An identical experiment in old animals was performed simultaneously (<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030525#pmed-0030525-t001" target="_blank">Table 1</a>, experiment 6), showed results indistinguishable from those of young mice (unpublished data). All tissues were stained with hematoxylin and eosin.</p

VRP-S Induces Short-Term Protection against icGDO3-S in Young and Partial Protection in Old Mice

<div><p>(A) Lung titers of BALB/c mice vaccinated and boosted with 10⁶ IU of VRP-S, VRP-N, a combination of VRP-S plus VRP-N (VRP-S+N), or mock vaccinated with PBS, then challenged with 10⁵ pfu of icGDO3-S challenge (<i>n</i> = 8 per group). Lungs were harvested 2 d post-challenge.</p> <p>(B) Lung titers of aged BALB/c mice vaccinated at greater than 26 wk of age, boosted 4 wk later, then challenged 12 wk post-boost with icGDO3-S (<i>n</i> = 7 VRP-N, <i>n</i> = 8 for other groups). Tissue was harvested 4 d post-challenge.</p> <p>(C) SARS CoV specific in situ signal (black arrows) was observed in the lungs of senescent mice that were vaccinated with PBS (unpublished data) or VRP-N (a) and challenged with icGDO3-S, although overall, the signal appeared to be less intense than that observed in the icSARS challenge animals (<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.0030525#pmed-0030525-g002" target="_blank">Figure 2</a>C). Vaccination with VRP-S (b) or VRP-S+N (c) failed to induce complete protection from icGD03-S challenge, as sections from two of five S- and three of four S+N-vaccinated animals exhibited signal above that of uninfected controls.</p></div