Table_1_Associations between prefrontal PI (16:0/20:4) lipid, TNC mRNA, and APOA1 protein in schizophrenia: A trans-omics analysis in post-mortem brain.xlsx

BackgroundThough various mechanisms have been proposed for the pathophysiology of schizophrenia, the full extent of these mechanisms remains unclear, and little is known about the relationships among them. We carried out trans-omics analyses by comparing the results of the previously reported lipidomics, transcriptomics, and proteomics analyses; all of these studies used common post-mortem brain samples.MethodsWe collected the data from three aforementioned omics studies on 6 common post-mortem samples (3 schizophrenia patients and 3 controls), and analyzed them as a whole group sample. Three correlation analyses were performed for each of the two of three omics studies in these samples. In order to discuss the strength of the correlations in a limited sample size, the p-values of each correlation coefficient were confirmed using the Student’s t-test. In addition, partial correlation analysis was also performed for some correlations, to verify the strength of the impact of each factor on the correlations.ResultsThe following three factors were strongly correlated with each other: the lipid level of phosphatidylinositol (PI) (16:0/20:4), the amount of TNC mRNA, and the quantitative signal intensity of APOA1 protein. PI (16:0/20:4) and TNC showed a positive correlation, while PI (16:0/20:4) and APOA1, and TNC and APOA1 showed negative correlations. All of these correlations reached at p ConclusionThe current results suggest that these three factors may provide new clues to elucidate the relationships among the candidate mechanisms of schizophrenia, and support the potential of trans-omics analyses as a new analytical method.</p

Data_Sheet_1_Associations between prefrontal PI (16:0/20:4) lipid, TNC mRNA, and APOA1 protein in schizophrenia: A trans-omics analysis in post-mortem brain.docx

BackgroundThough various mechanisms have been proposed for the pathophysiology of schizophrenia, the full extent of these mechanisms remains unclear, and little is known about the relationships among them. We carried out trans-omics analyses by comparing the results of the previously reported lipidomics, transcriptomics, and proteomics analyses; all of these studies used common post-mortem brain samples.MethodsWe collected the data from three aforementioned omics studies on 6 common post-mortem samples (3 schizophrenia patients and 3 controls), and analyzed them as a whole group sample. Three correlation analyses were performed for each of the two of three omics studies in these samples. In order to discuss the strength of the correlations in a limited sample size, the p-values of each correlation coefficient were confirmed using the Student’s t-test. In addition, partial correlation analysis was also performed for some correlations, to verify the strength of the impact of each factor on the correlations.ResultsThe following three factors were strongly correlated with each other: the lipid level of phosphatidylinositol (PI) (16:0/20:4), the amount of TNC mRNA, and the quantitative signal intensity of APOA1 protein. PI (16:0/20:4) and TNC showed a positive correlation, while PI (16:0/20:4) and APOA1, and TNC and APOA1 showed negative correlations. All of these correlations reached at p ConclusionThe current results suggest that these three factors may provide new clues to elucidate the relationships among the candidate mechanisms of schizophrenia, and support the potential of trans-omics analyses as a new analytical method.</p

Decrease in Sphingomyelin (d18:1/16:0) in Stem Villi and Phosphatidylcholine (16:0/20:4) in Terminal Villi of Human Term Placentas with Pathohistological Maternal Malperfusion

<div><p>Placental villi play pivotal roles in feto-maternal transportation and phospholipids constitute a major part of the villous membrane. We have been developing and optimizing an imaging system based on a matrix-assisted laser desorption/ionization (MALDI)-based mass spectrometer, which provides clear two-dimensional molecular distribution patterns using highly sensitive mass spectrometry from mixtures of ions generated on tissue surfaces. We recently applied this technology to normal human uncomplicated term placentas and detected the specific distribution of sphingomyelin (SM) (d18:1/16:0) in stem villi and phosphatidylcholine (PC) (16:0/20:4) in terminal villi. In the present study, we applied this technology to nine placentas with maternal or fetal complications, and determined whether a relationship existed between these specific distribution patterns of phospholipid molecules and the six representative pathological findings of placentas, i.e., villitis of unknown etiology (VUE), thrombus, atherosis, chorioamnionitis (CAM), immature terminal villi, and multiple branched terminal villi. In two placentas with the first and second largest total number of positive pathological findings, i.e., five and three positive findings, the specific distribution of SM (d18:1/16:0) in stem villi and PC (16:0/20:4) in terminal villi disappeared. The common pathological findings in these two placentas were atherosis, immature terminal villi, and multiple branched terminal villi, suggesting the possible involvement of the underperfusion of maternal blood into the intervillous space. On the other hand, the number of pathological findings were two or less in the seven other placentas, in which no specific relationships were observed between the differential expression patterns of these two phospholipids in stem and terminal villi and the pathological findings of the placentas; however, the specific distribution pattern of SM (d18:1/16:0) in stem villi disappeared in four placentas, while that of PC (16:0/20:4) in terminal villi was preserved. These results suggested that the absence of the specific distribution of PC (16:0/20:4) in terminal villi, possibly in combination with the absence of SM (d18:1/16:0) in stem villi, was linked to placental morphological changes in response to maternal underperfusion of the placenta.</p></div

Expression profiles of sphingomyelin (d18:1/16:0) in stem villi and phosphatidylcholine (16:0/20:4) in terminal villi by imaging mass spectrometry.

<p>Samples grouped as—showed significant decreases (p<0.05) with an analysis of variance and Tukey’s test from the other group denoted as +.</p><p>Expression profiles of sphingomyelin (d18:1/16:0) in stem villi and phosphatidylcholine (16:0/20:4) in terminal villi by imaging mass spectrometry.</p

Representative averaged mass spectra from entire sections.

<p>Signals collected between <i>m/z</i> 700–900 were shown for sample Nos.2 (A), 8 (B), and 9 (C) in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t003" target="_blank">Table 3</a>. Peaks corresponding to representative phospholipids were labeled. Imaging results for <i>m/z</i> 725.5 and <i>m/z</i> 804.5 were shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.g003" target="_blank">Fig 3</a> and summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t003" target="_blank">Table 3</a>. Pathological findings of the placenta were summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t001" target="_blank">Table 1</a>.</p

Ion images for <i>m/z</i> 725.5 (B, F, J), <i>m/z</i> 804.5 (C, G, K), and <i>m/z</i> 734.5 (D, H, L).

<p>A-D; placenta No. 2. E-H; placenta No.8. I-L; placenta No. 9. The peaks of <i>m/z</i> 725.5 corresponding to sphingomyelin (d18:1/16:0) and <i>m/z</i> 804.5 corresponding to phosphatidylcholine (16:0/20:4) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.ref021" target="_blank">21</a>] were visualized. Imaging results for <i>m/z</i> 725.5 and <i>m/z</i> 804.5 associated with pathological findings were summarized in Tables <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t001" target="_blank">1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t003" target="_blank">3</a>. An ion image of <i>m/z</i> 734.5 (D, H, L) was presented as a positive control independent of pathological findings. A, E, I; HE staining of consecutive sections. Pathological findings of the placenta were summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142609#pone.0142609.t001" target="_blank">Table 1</a>. The white arrows indicate stem villi. The areas surrounded by white dotted lines correspond to stem villi, in which the specific distribution of <i>m/z</i> 725.5 was observed in (J), but not in (B) or (F). The outside areas of white dotted circles are mostly terminal villi, in which the specific distribution of <i>m/z</i> 804.5 was observed in (G) and (K), but not in (C). +; Preservation of the specific distribution of <i>m/z</i> 734.5 in stem villi (J) or <i>m/z</i> 804.5 in terminal villi (G, K). Red squares also indicate the preservation of the specific distribution of <i>m/z</i> 734.5 in stem villi (J) or <i>m/z</i> 804.5 in terminal villi (G, K). −; Absence of the specific distribution of <i>m/z</i> 734.5 in stem villi (B, F) or <i>m/z</i> 804.5 in terminal villi (C).</p

Pathological findings in placentas enrolled.

<p>VUE; villitis of unknown etiology. CAM; Chorioamnionitis.</p><p>Pathological findings in placentas enrolled.</p

Skin structure and lipid composition of skin.

<p>Upper panel shows a model of the structure of the skin. Lower panel shows the results of thin-layer chromatographic analyses of lipid in mouse skin. We assessed lipid composition enriched in mouse footpad skin by TLC analysis. Cer and PC are shown as major lipid species. In addition we also detected GlcCer (a), SM, and TAG (b).The arrowhead shows the localization of standard, Cer (d18∶1/C24∶0) and PC (C16∶0/C16∶0).</p

Testosterone inhibits oxidative stress-induced endothelial senescence through eNOS/SIRT1.

<p><b>A.</b> Testosterone inhibited SA-βgal activity and senescent morphological appearance induced by hydrogen peroxide (100 µmol/L). <b>B.</b> Expression of eNOS, SIRT1, and PAI-1 in hydrogen peroxide (100 µmol/L)-treated HUVEC under treatment with DHT or testosterone. <b>C.</b> Overexpression of SIRT1 and DHT reduced SA-βgal activity. eNOS expression was increased by overexpression of SIRT1, and DHT increased phosphorylation of eNOS (Ser1177). <b>D.</b> SIRT1 inhibition by siRNA or sirtinol (100 µmol/L) abrogated the effect of testosterone on SA-βgal activity. <b>E.</b> Treatment with testosterone or DHT increased eNOS activity. <b>F.</b> eNOS inhibition by siRNA or L-NAME (10 mM) abrogated the effect of testosterone on SA-βgal activity. <b>G.</b> Treatment with L-NAME decreased SIRT1 expression in DHT-treated HUVEC. (*p<0.05, N = 3).</p

Supplementation of testosterone improves cognitive function in SAMP8 mice.

<p><b>A.</b> Escape latency and plasma testosterone level of male SAMR1 (N = 10) and SAMP8 mice (N = 10) at 18 months of age. These mice were implanted subcutaneously with a placebo or a 21-day-release 2.5 mg testosterone pellet in the dorsal neck. <b>B.</b> Number of SA-βgal-stained Leydig cells in testes in SAMR1 and SAMP8. Arrows indicate Leydig cells. Representative SA-βgal-stained testes from SAMR1 and SAMP8. <b>C.</b> Escape latency of castrated SAMR1 (upper, N = 5) and recipient SAMP8 (lower, N = 5). Observation (0–10 weeks) was started from 3 weeks after operation. <b>D.</b> SIRT1 expression in hippocampus of SAMP8 with or without DHT treatment. Immunofluorescent staining for SIRT1 (green) and DAPI (blue). <b>E.</b> Expression of AR in SAMR1 and SAMP8 brains. (*p<0.05).</p