Additional file 6: Figure S5. of Insertionally polymorphic sites of human endogenous retrovirus-K (HML-2) with long target site duplications

Model of pseudo-TSDs flanking a solo LTR. This model may explain the long homologous sequences flanking solo LTRs. In this model, a plausible preintegration site is formed via homologous recombination between genomic regions containing a solo LTR within two segmentally duplicated regions (locus A and locus B). This model may also apply to pseudo-TSDs flanking a provirus. Green and red areas are TSD sequences for proviral integration at locus A and locus B, respectively. (PDF 139 kb

Additional file 1: Table S1. of Insertionally polymorphic sites of human endogenous retrovirus-K (HML-2) with long target site duplications

Insertional polymorphisms in HML-2; Table S2. Source information for DGV data; Table S3. Primer information; Table S4. Long putative TSD sequences flanking LTR5_Hs; Table S5. Putative sites for HML-2 insertional polymorphisms. (XLSX 27 kb

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

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

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

Senescent endothelial cells of hippocampus are decreased by treatment with DHT.

<p><b>A.</b> Oxidative stress level was measured by detection of carbonyl groups introduced into proteins. <b>B.</b> Acetyl-choline concentration was measured by a colorimetric method. <b>C.</b> SA-βgal-stained endothelial cells and SIRT1 expression in CA3 area of hippocampus in SAMR1 and SAMP8 with or without DHT treatment. Immunofluorescent staining for SIRT1 (green), PECAM-1 (red), and DAPI (blue). <b>D.</b> Number of SA-βgal-stained endothelial cells in CA3 area of hippocampus in SAMR1 and SAMP8 with or without DHT treatment. <b>E.</b> Expression of SIRT1, PECAM-1, and β-actin was analyzed using cerebral micro vascular cells. <b>F.</b> Escape latency of SAMR1 (N = 10) and SAMP8 mice (N = 10). Male mice were treated daily for 2 weeks with DHT (500 µg s.c) and L-NAME (20 mg/kg gavage) before trials. <b>G.</b> Escape latency of SAMR1 (N = 5) and SAMP8 mice (N = 5). Male mice were treated daily for 2 weeks with DHT (500 µg s.c) and L-VNIO (5 mg/kg IP) before trials. (*p<0.05, n.s: not significant).</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

Testosterone deficiency causes senescence of hippocampus and cognitive impairment in SAMP8 mice.

<p><b>A.</b> Body weight, appearance, and plasma testosterone level of male SAMR1 and SAMP8 mice at 12 weeks of age. <b>B.</b> Escape latency of SAMR1 (N = 10) and SAMP8 mice (N = 10). Male mice were treated daily for 2 weeks with DHT (500 µg s.c) before trials. Swim speed during quadrant test on day 10. <b>C.</b> Total distance and the ratio of central/total distance were measured in open field tests. <b>D.</b> Number of amyloid ß plaques, pyramidal cells, and SA-ßgal-positive cells in CA1 and CA3 areas of hippocampus in SAMR1 and SAMP8. (*p<0.05, n.s: not significant).</p

Oxidative stressed-induced endothelial cell senescence promotes adjacent neuronal cell senescence.

<p><b>A.</b> Co-culture cell culture dish. <b>B.</b> Number of SA-βgal-stained MHC and senescent appearance of MHC were increased, and acetyl-choline concentration was decreased by co-culture with senescent endothelial cells. Senescent MHC are indicated by arrows. <b>C.</b> Expression of SIRT1, PAI-1, p53, and β-actin in MHC co-cultured with senescent endothelial cells. <b>D.</b> Expression of IL-6, IL-8, MCP-1, and TNF-α in endothelial cells were analyzed by RT-PCR. <b>E.</b> The number of SA-βgal-stained MHC was decreased by treatment with testosterone in both MHC and HUVEC (MHC, testosterone (+)), or HUVEC (MHC, testosterone (−)) alone. <b>F.</b> Resveratrol decreased the number of SA-βgal-stained MHC co-cultured with senescent endothelial cells. (*p<0.05, N = 3). <b>G.</b> Hypothetical signal transduction pathways of testosterone in endothelial cells.</p

Detection of Distinct Distributions of Acetaminophen and Acetaminophen-Cysteine in Kidneys up to 10 μm Resolution and Identification of a Novel Acetaminophen Metabolite Using an AP-MALDI Imaging Mass Microscope

Drug distribution studies in tissue are crucial for understanding the pharmacokinetics and potential toxicity of drugs. Recently, matrix-assisted laser desorption ionization-mass spectrometry imaging (MALDI-MSI) has gained attention for drug distribution studies due to its high sensitivity, label-free nature, and ability to distinguish between parent drugs, their metabolites, and endogenous molecules. Despite these advantages, achieving high spatial resolution in drug imaging is challenging. Importantly, many drugs and metabolites are rarely detectable by conventional vacuum MALDI-MSI because of their poor ionization efficiency. It has been reported that acetaminophen (APAP) and one of its major metabolites, APAP-Cysteine (APAP-CYS), cannot be detected by vacuum MALDI-MSI without derivatization. In this context, we showed the distribution of both APAP and APAP-CYS in kidneys at high spatial resolution (25 and 10 μm) by employing an atmospheric pressure-MALDI imaging mass microscope without derivatization. APAP was highly accumulated in the renal pelvis 1 h after drug administration, while APAP-CYS exhibited characteristic distributions in the outer medulla and renal pelvis at both 30 min and 1 h after administration. Interestingly, cluster-like distributions of APAP and APAP-CYS were observed in the renal pelvis at 10 μm spatial resolution. Additionally, a novel APAP metabolite, tentatively coined as APAP-butyl sulfate (APAP-BS), was identified in the kidney, brain, and liver by combining MSI and tandem MSI. For the first time, our study revealed differential distributions of APAP, APAP-CYS (in kidneys), and APAP-BS (in kidney, brain, and liver) and is believed to enhance the understanding of the pharmacokinetics and potential nephrotoxicity of this drug