Experimental cohorts: number of animals per cohort.

<p>*Published <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088916#pone.0088916-Ormond1" target="_blank">[14]</a>.</p

Stem cell therapy in conjunction with curcumin synergistically improves recovery from moderate or severe SCI, as demonstrated by improved body weight gain, which correlated with BBB score.

<p><b>A:</b> Stem cell transplanted animals showed improved body weight gain in comparison to combinational treatment or DMSO treated controls. <b>B:</b> A positive correlation exists between body weight gain and improved BBB score in moderate SCI. R² values are listed next to their respective cohort. <b>C:</b> Combination of NSC therapy with curcumin treatment showed significant improvement in body weight gain in comparison to DMSO control. Curcumin treated rats showed a trend toward significance in BW gain. <b>D:</b> Correlation of BBB scores with body weight after severe SCI. R² values are listed next to their respective cohort.</p

Histopathological examination after severe SCI.

<p><b>A:</b> NSC plus curcumin and NSC all displayed enhanced spared area relative to the total area suggesting an improvement in recovery. DMSO treated rats also demonstrated increased spared area of the spinal cord, but somewhat less than the NSC plus curcumin, or NSC treatments. Curcumin-treated rats demonstrated less spared area than the other three treatments. <b>B:</b> Photomicrographs of representative samples of spinal cord sections across the area of contusion stained with Lox blue.</p

Curcumin enhances stem cell proliferation of NSC derived from SVZ in low doses, however it induces apoptosis at high doses.

<p><b>A:</b> Schematic presentation of treatments timeline used in moderate and severe SCI. <b>B:</b> Micropunched area of the subventricular zone (SVZ), depicting the presence of ependymal cells (brown), astrocytes (blue), transit amplifying cells (green) and neuroblasts (red). <b>C:</b> Immunofluorescence analysis demonstrates the presence of stem cell marker nanog in neurospheres and DAPI staining for DNA. <b>D, E:</b> Effect of curcumin on neurosphere formation. At a low concentration of 500 nM, curcumin treatment stimulated proliferation of NSC in 24 hr. At higher concentrations (1 uM), curcumin caused autolysis and neurosphere fragmentation. The chemical structure of curcumin is also displayed.</p

Soleus muscle weight as an indicator of recovery after moderate and severe SCI, and correlated with BBB score.

<p><b>A:</b> Rats with greater recovery of hindlimb motor function have better soleus muscle weight at necropsy. While stem cell transplanted animals and combinational treatments had improved soleus muscle weight in comparison to DMSO controls, this did not reach statistical significance. <b>B:</b> Correlation of BBB score with soleus muscle weight after moderate SCI demonstrates strong correlation (0.97). Values presented are mean ± SEM. *p<0.05. <b>C:</b> Soleus muscle weight after severe SCI showed that all cohorts had larger soleus muscle mass compared to DMSO treated controls, however curcumin or stem cell monotherapy showed only a trend towards significance. NSC plus curcumin treatment improved soleus muscle weight significantly in comparison to DMSO controls. <b>D:</b> Correlation of BBB score with soleus muscle weight after severe SCI demonstrated a strong correlation (r² = 0.91). Values presented are mean ± SEM. *p<0.05.</p

Stem cell therapy in conjunction with low-dose curcumin had a synergistic effect in recovery from severe SCI, as demonstrated by improved BBB score.

<p><b>A:</b> Functional analysis (BBB) after moderate SCI showed that stem cell therapy with a high dose of curcumin rendered BBB scores, which were statistically significant as compared to control rats (DMSO), starting from week 3. <b>B:</b> In severe SCI, rats undergoing combined NSC transplantation and curcumin showed consistent improvement in BBB score as compared to DMSO treated controls. Statistical analysis of combined NSC+curcumin treatment also demonstrated significant improvement in comparison to NSC or curcumin treatment alone.</p

Identification and Characterization of Differentially Expressed Genes in Inferior and Superior Spikelets of Rice Cultivars with Contrasting Panicle-Compactness and Grain-Filling Properties

<div><p>Breeding programs for increasing spikelet number in rice have resulted in compactness of the panicle, accompanied by poor grain filling in inferior spikelets. Although the inefficient utilization of assimilate has been indicated as responsible for this poor grain filling, the underlying cause remains elusive. The current study utilized the suppression subtractive hybridization technique to identify 57 and 79 genes that overexpressed in the superior and inferior spikelets (with respect to each other), respectively, of the compact-panicle rice cultivar Mahalaxmi. Functional categorization of these differentially expressed genes revealed a marked metabolic difference between the spikelets according to their spatial location on the panicle. The expression of genes encoding seed storage proteins was dominant in inferior spikelets, whereas genes encoding regulatory proteins, such as serine-threonine kinase, zinc finger protein and E3 ligase, were highly expressed in superior spikelets. The expression patterns of these genes in the inferior and superior spikelets of Mahalaxmi were similar to those observed in another compact-panicle cultivar, OR-1918, but differed from those obtained in two lax-panicle cultivars, Upahar and Lalat. The results first suggest that the regulatory proteins abundantly expressed in the superior spikelets of compact-panicle cultivars and in both the superior and inferior spikelets of lax-panicle cultivars but poorly expressed in the inferior spikelets of compact-panicle cultivars promote grain filling. Second, the high expression of seed-storage proteins observed in the inferior spikelets of compact-panicle cultivars appears to inhibit the grain filling process. Third, the low expression of enzymes of the Krebs cycle in inferior spikelets compared with superior spikelets of compact-panicle cultivars is bound to lead to poor ATP generation in the former and consequently limit starch biosynthesis, an ATP-consuming process, resulting in poor grain filling.</p></div

Stat-5A transfection confers resistance to CD4⁺ T cells from tumor-induced death.

<p>A, Stat-5A (<i>left panel</i>) and Stat-5B (<i>right panel</i>) isoforms were immunoprecipitated from cell lysates using specific antibodies and then Western blotted with anti-phospho-tyrosine or anti-Stat-5A/Stat-5B antibodies to determine phosphorylation status of specific proteins. B, Jurkat T cells were transfected with control vector, wild-type <i>Stat-5A/Stat-5B</i>, C-terminal truncated <i>Stat-5A</i>₇₁₃/<i>Stat-5B</i>₇₁₈ or constitutively active <i>Stat-5A1*6</i> genes and were cultured in the presence of media alone or MCF-7 spent media (±theaflavins) for 48 h. Percent cell death (Annexin-V-PE⁺/7AAD⁺) was determined flow cytometrically. Values are mean±S.E.M. of three independent sets of experiments.</p

Re-confirmation of PGE₂ as the molecule behind tumor-induced perturbation in CD4⁺ T cell survival signaling.

<p>A, MCF-7 cells were treated with theaflavins or celecoxib or transfected with Cox-2-siRNA and the levels of Cox-2 and GAPDH (internal control) mRNA were determined by RT-PCR (<i>upper panel</i>). Western blot analysis was performed for the determination of levels of Cox-2 or α-Actin (internal control) proteins (<i>middle panel</i>). In parallel experiments the amount of tumor-secreted PGE₂ in the cell-free supernatant was determined by ELISA (<i>lower panel</i>). B, Purified CD4⁺ T cells were cultured in the presence of media alone or MCF-7-spent media (tumors were either pre-treated with 25 µg/ml theaflavins/3.5 ng/ml PGE₂/50 µM celecoxib or transfected with 300pmole Cox-2-siRNA) for 48 h. Expression levels of IL2Rγc and Bcl-2 as well as phosphorylation status of Jak-3/-Stat-5 were determined by Western blotting in which α-Actin was used as internal control (<i>upper panel</i>). In parallel experiments, flowcytometric determination of percent cell death (<i>lower panel</i>) was established. Values are mean±S.E.M. of three independent experiments.</p

Tumor-shed PGE₂ is responsible for CD4⁺ T cell apoptosis.

<p>A, Tumor-secreted PGE₂ over time in cell-free spent media of MCF-7 cells (control (○), Cox-2-siRNA-transfected (Δ) or theaflavin-treated (•) was determined by ELISA. B, Percent CD4⁺ T cell death (Annexin-V-PE⁺/7AAD⁺), induced by the spent media as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007382#pone-0007382-g002" target="_blank">Fig. 2A</a>, was plotted over time. Values are mean±S.E.M. of three independent sets of experiments.</p