Microglial p38β MAPK deficiency failed to protect cortical neurons against LPS-induced neurotoxicity in microglia/neuron co-culture.

<p>WT mouse primary cortical neurons were plated on coverslips at 5×10⁴/well, and were co-cultured with microglia (2×10⁴/well) from WT or p38β KO mice. Cells were treated with either vehicle or LPS (3 ng/ml) for 72 hr, followed by trypan blue exclusion assay to evaluate neuronal survival. LPS treatment induced significant neuronal death in WT microglia/WT neuron co-culture. Similar levels of neuronal death were seen in LPS-treated co-cultures of p38β KO microglia/WT neurons. Data represent the mean ± SEM from 2–4 independent experiments. ****p<0.0001 WT-veh vs. WT-LPS; ^####p<0.0001 KO-veh vs. KO-LPS.</p

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-6

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>ignificantly induced NO generation, and pretreatment with pioglitazone inhibited this LPS-induced NO production in a dose-dependent manner. Data presented are representative of three independent experiments (n = 3). (***< 0.001 vs. control ###< 0.001 vs. LPS

Verification of p38β KO in microglia and brain.

<p>Primary microglia from mouse cortex were prepared as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056852#s2" target="_blank">Methods</a> and plated at 1×10⁴ cells/well in 48 well plates. Total RNA from microglia cultures (<b>A</b>) or from mouse cortical tissue (<b>B</b>) derived from WT (white bars) or p38β KO (black bars) mice was isolated, and the mRNA levels of different p38 MAPK isoforms were determined by qPCR. In both the microglia cultures and the brain tissues, p38β mRNA was readily measureable in WT mice but was not detected in the p38β KO mice. The p38α MAPK isoform in both microglia and cortex was expressed at much higher levels than any of the other isoforms, and there was no significant difference between the levels of p38α in either WT or p38β KO mice. The levels of p38δ mRNA were very low to undetectable in both WT and p38β KO mice. The expression of p38γ mRNA was slightly higher in microglia cultures from p38β KO mice compared to microglia from WT mice, but this difference was not seen in the cortical tissue samples. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056852#s3" target="_blank">Results</a> are expressed as fold change compared to p38β levels, and represent the mean ± SEM of two determinations.</p

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-1

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>ent with pioglitazone (10 μM), 1 hr before LPS, prevents its expression. : Rat mesencephalic mixed cultures were treated with the selective iNOS inhibitor 1400 W, with different doses from 1 ng/ml to 10 μM/ml, 1 hr before a 72 hr LPS exposure. The number of TH-positive neurons was determined by immunocytochemistry. Data presented are representative of three independent experiments (n = 3). (**< 0.01 vs. control, ***< 0.001 vs. control, #< 0.05 vs. LPS, ##< 0.01 vs. LPS, ###< 0.001 vs. LPS)

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-4

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>tivation was assessed after 10 min, and P13K and Akt were assessed after 60 min. PPAR-γ activation, PI3K and Akt expression were observed in the pioglitazone-treated cultures, compared to control and LPS-only groups. Data presented are representative of three independent experiments (n = 3). (*< 0.05 vs. control, #< 0.05 vs. LPS, ##< 0.01 vs. LPS)

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-0

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>ignificantly induced NO generation, and pretreatment with pioglitazone inhibited this LPS-induced NO production in a dose-dependent manner. Data presented are representative of three independent experiments (n = 3). (***< 0.001 vs. control ###< 0.001 vs. LPS

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-5

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>azone followed by LPS 60 mins later in microglia-enriched culture, and after 48 hrs NO levels were measured. The results show that the LPS-induced NO level was significantly higher than control (< 0.01), and that pretreatment with pioglitazone inhibits LPS-induced NO (< 0.01). In contrast, pretreatment with wortmannin enhanced the LPS-induced increase in NO generation (< 0.05), and this pretreatment prevented the inhibitory effect of pioglitazone on LPS-induced NO generation. Data presented are representative of three independent experiments (n = 3).). (**< 0.01 vs. control, #< 0.05 vs. LPS, ##< 0.01 vs. LPS, &&< 0.01 vs. Piog plus LPS)

Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt-2

<p><b>Copyright information:</b></p><p>Taken from "Pioglitazone inhibition of lipopolysaccharide-induced nitric oxide synthase is associated with altered activity of p38 MAP kinase and PI3K/Akt"</p><p>Journal of Neuroinflammation 2008;5():4-4.</p><p>Published online 18 Jan 2008</p><p>PMCID:PMC2254610.</p><p></p>(1 μg/ml) exposure and, after 24 hrs, NO levels were measured. Only the p38 MAPK inhibitor prevented NO production. : Pretreatment with pioglitazone inhibited LPS-induced phosphorylation of p38 MAPK in mesencephalic neuronal-microglia mixed cultures. Pioglitazone was added 1 hr before LPS treatment (1 μg/ml) and, after 30 mins, p38 MAPK was immunobloted. As shown in 3B, LPS increased phosphorylation of p38 MAPK, and pretreatment with pioglitazone inhibited this expression. Data presented are representative of three independent experiments (n = 3). (*< 0.05 vs. control, ***< 0.001 vs. control, #< 0.05 vs. LPS)

Correlations of Pituitary Tumor Transforming Gene Expression with Human Pituitary Adenomas: A Meta-Analysis

<div><p>Objective</p><p>Pituitary tumor transforming gene (PTTG) is an important paracrine growth factor involved in early lactotrope transformation and early onset of angiogenesis in pituitary hyperplasia. Emerging evidences have shown that PTTG expression may contribute to the etiology of pituitary adenomas; but individually published studies showed inconclusive results. This meta-analysis aimed to derive a more precise estimation of the correlations of PTTG expression with human pituitary adenomas.</p><p>Methods</p><p>A range of electronic databases were searched: MEDLINE (1966∼2013), the Cochrane Library Database (Issue 12, 2013), EMBASE (1980∼2013), CINAHL (1982∼2013), Web of Science (1945∼2013) and the Chinese Biomedical Database (CBM) (1982∼2013) without language restrictions. Meta-analysis was performed using the STATA 12.0 software. Crude odds ratio (OR) or standard mean difference (SMD) with its corresponding 95% confidence interval (95%CI) were calculated.</p><p>Results</p><p>Twenty-four clinical cohort studies were included with a total of 1,464 pituitary adenomas patients. The meta-analysis results revealed that patients with invasive pituitary adenomas had higher positive expression of PTTG than those of non-invasive patients (OR  = 6.68, 95%CI  = 3.72–11.99, <i>P</i><0.001). We also found a significant difference in microvessel density between invasive and non-invasive patients (SMD  = 1.81, 95%CI  = 0.39–3.23, <i>P</i> = 0.013). However, there were no significant difference in PTTG expression between functional and non-functional patients with pituitary adenomas (OR  = 1.11, 95%CI  = 0.58–2.10, <i>P</i> = 0.753). No publication bias was detected in this meta-analysis (all <i>P</i>>0.05).</p><p>Conclusion</p><p>This present meta-analysis suggests that PTTG expression may be associated with tumor invasiveness and microvessel density of pituitary adenomas, while no correlations with functional status was found.</p></div

Forest plots for the relationship between PTTG expression and tumor invasiveness of pituitary adenomas.

<p>Forest plots for the relationship between PTTG expression and tumor invasiveness of pituitary adenomas.</p