Targeting prolyl-isomerase Pin1 prevents mitochondrial oxidative stress and vascular dysfunction: insights in patients with diabetes

The present study demonstrates that Pin1 is a common activator of key pathways involved in diabetic vascular disease in different experimental settings including primary human endothelial cells, knockout mice, and diabetic patients. Gene silencing and genetic disruption of Pin1 prevent hyperglycaemia-induced mitochondrial oxidative stress, endothelial dysfunction, and vascular inflammation. Moreover, we have translated our findings to diabetic patients. In line with our experimental observations, Pin1 up-regulation is associated with impaired flow-mediated dilation, increased oxidative stress, and plasma levels of adhesion molecules. In perspective, these findings may provide the rationale for mechanism-based therapeutic strategies in patients with diabete

Motoric Cognitive Risk Syndrome, Subtypes and 8-Year All-Cause Mortality in Aging Phenotypes: The Salus in Apulia Study

BackgroundThis study aims to establish the key clinical features of different motoric cognitive risk (MCR) subtypes based on individual quantitative measures of cognitive impairment and to compare their predictive power on survival over an 8-year observation time.MethodsWe analyzed data from a population-based study of 1138 subjects aged 65 years and older in south Italy. These individuals were targeted and allocated to subtypes of the MCR phenotype according to the slowness criterion plus one other different cognitive domain for each characterized phenotype (Subjective Cognitive Complaint [SCC]; Global Function [Mini Mental State Examination (MMSE) < 24]; or a combination of both). Clinical evaluation and laboratory assays, along with a comprehensive battery of neuropsychological and physical tests, completed the sample investigation.ResultsMCR prevalence was found to be 9.8% (n = 112), 3.6% (n = 41), 3.4% (n = 39) and 1.8% (n = 21) for the MCR, MCR-GlobalFunction, MCR-StructuredSCC and MCR-SCC and GlobalFunction, respectively. Univariate Cox survival analysis showed an association only of the MCR-GlobalFunction subtype with an almost three-fold increased risk of overall death as compared to the other counterparts (HR 2.53, 95%CI 1.28 to 4.99) over an 8-year observation period. Using Generalized Estimating Equations (GEE) for clustered survival data, we found that MCR males had an increased and significant mortality risk with respect to MCR female subjects.ConclusionsMCR phenotypes assigned to the MMSE cognitive domain are more likely to have an increased risk of overall mortality, and gender showed a huge effect on the risk of death for MCR subjects over the 8-year observation

Serum levels of IL-6 are associated with cognitive impairment in the salus in apulia population-based study

Growing evidence suggests that inflammation contributes to brain aging and neurodegeneration. This study investigates the relationship between global cognitive as well executive function and the inflammatory markers IL-6, CRP, and TNF-α in a population-based study of older adults. A population-based sample, of older people in Southern Italy, was enrolled. We measured serum levels of IL-6, CRP, and TNF-α. We also administered two neuropsychological tests: Mini-Mental State Examination and Frontal Assessment Battery. Rank-based regression models were performed to investigate the relationship between inflammatory markers and cognitive functions, including major demographic and clinical confounders for adjustment. The sample consisted of 1929 subjects aged between 65 and 95 years. Multivariate linear regression analysis revealed that higher serum levels of IL-6 were associated with lower MMSE and FAB scores even after adjustment for demographic data and cardiovascular risk factors. No significant associations were found between cognitive functioning and serum levels of CRP and TNF-α. Our results suggest that higher levels of IL-6 were associated with cognitive impairment in an older adult population of Southern Italy

Intensive Neurofeedback-based Training to Improve Impaired Attention and Executive Functions Secondary to Resection of Tuberculum Sellae Meningioma: A Case Study

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Targeting Prolyl-isomerase-1 protects against hyperglycemia-induced endothelial dysfunction and vascular inflammation: alterations in patients with type 2 diabetes

Introduction: Prolyl-isomerase-1 (Pin1) regulates function of protein substrates through isomerization of peptide bonds that link phosphoserine or phosphothreonine to proline. Recent studies showed that Pin1 favours cancerogenesis through reactive oxygen species (ROS) production and inflammation. Whether Pin1 partecipates to vascular disease is unknown. Hypothesis: This study investigates the role of Pin1 in diabetes-related vascular dysfunction. Methods: Human aortic endothelial cells (HAECs) were exposed to normal (NG, 5mmol/L) or high glucose concentrations (HG, 25 mmol/L) in the presence or in the absence of Pin1 pharmacological inhibitor Juglone or siRNA-mediated knockdown. Diabetes was induced by streptozotocin in C57/B6 mice and animals treated with Pin1 siRNA i.v or Juglone i.p for 30 days. Endothelial function was assessed by dose-response curve with acetylcoline (10-9-10-6 mol/L). Mitochondrial ROS were measured by ESR spectroscopy. Pin1 gene expression was assessed in peripheral blood monocytes (PBM) of 37 patients with type 2 diabetes (T2DM) and 20 age-matched controls and correlated with flow-mediated vasodilation (FMD) of the brachial artery, urinary oxidative marker 8-isoprostaglandinF2α (8-isoPGF2α) and plasma adhesion molecules VCAM-1, ICAM-1 and MCP-1. Results: Pin1 expression increased in HAECs exposed to HG (289±22% vs. NG, p<0.01) and aortas of diabetic mice (216±32% vs. controls, p<0.05). Immunoprecipitation showed that Pin1 recognizes phosphoserine motifs of the pro-oxidant mitochondrial adaptor p66Shc and NF-kB p65. Interestingly, Juglone or Pin1siRNA prevented p66Shc-induced ROS production and suppressed NF-kB-dependent upregulation of adhesion molecules. In diabetic mice, silencing of Pin1 or Juglone prevented endothelial dysfunction, ROS production and vascular inflammation. Of note, Pin1 mRNA was significantly upregulated in PBM of T2DM patients as compared with controls (370±97 vs. 25±28,p<0.01) and correlated with FMD (r=-0.33, p<0.01), urinary 8-isoPGF2α (r=0.46, p<0.05), VCAM-1 (r=0.58, p<0.05) and ICAM-1 (r=0.55, p<0.05). Conclusions: Pin1 may critically participate to vascular disease in T2DM patients

Transcriptional Profiling of Hippocampus Identifies Network Alterations in Alzheimer's Disease

Alzheimer’s disease (AD) is a neurodegenerative disease characterized by rapid brain cell degeneration affecting different areas of the brain. Hippocampus is one of the earliest involved brain regions in the disease. Modern technologies based on high-throughput data have identified transcriptional profiling of several neurological diseases, including AD, for a better comprehension of genetic mechanisms of the disease. In this study, we investigated differentially expressed genes (DEGs) from six Gene Expression Omnibus (GEO) datasets of hippocampus of AD patients. The identified DEGs were submitted to Weighted correlation network analysis (WGCNA) and ClueGo to explore genes with a higher degree centrality and to comprehend their biological role. Subsequently, MCODE was used to identify subnetworks of interconnected DEGs. Our study found 40 down-regulated genes and 36 up-regulated genes as consensus DEGs. Analysis of the co-expression network revealed ACOT7, ATP8A2, CDC42, GAD1, GOT1, INA, NCALD, and WWTR1 to be genes with a higher degree centrality. ClueGO revealed the pathways that were mainly enriched, such as clathrin coat assembly, synaptic vesicle endocytosis, and DNA damage response signal transduction by p53 class mediator. In addition, we found a subnetwork of 12 interconnected genes (AMPH, CA10, CALY, NEFL, SNAP25, SNAP91, SNCB, STMN2, SV2B, SYN2, SYT1, and SYT13). Only CA10 and CALY are targets of known drugs while the others could be potential novel drug targets.</jats:p

Epigenetic remodeling of Nox2 by acetyltransferase Gcn5 drives hyperglycemia-induced endothelial oxidative stress

Introduction: Epigenetic modifications are recently emerging as important modulators of gene expression. The mammalian acetyltransferase gene non-derepressible 5 (GCN5) causes Nox2 upregulation in neutrophils through acetylation of histones. Whether GCN5 modulates oxidative stress in the cardiovascular system remains largely unknown. Hypothesis: The present study investigates GCN5 role in the modulation of diabetes-related endothelial oxidative stress. Methods: Human aortic endothelial cells (HAECs) were exposed to normal (NG, 5 mmol/L) or high glucose concentrations (HG, 25 mmol/L) in the presence or in the absence of GCN5 pharmacological inhibitor CPTH2 or siRNA-mediated knockdown. Superoxide anion (O2-) was measured by ESR spectroscopy. Chromatin immunoprecipitation (ChIP) was performed to investigate epigenetic modifications on Nox2 promoter induced by GCN5. In parallel, 30 patients with type 2 diabetes (T2DM) and 18 age-matched healthy controls were enrolled. All patients underwent flow-mediated vasodilation (FMD) of the brachial artery to assess endothelial function. Urinary levels of 8-isoprostaglandinF2α (8-isoPGF2α) were measured as a marker of oxidative stress. Gene expression of GCN5 and Nox2 was assessed from peripheral blood monocytes (PBM). Data are presented as fold change (FC). Results: HG caused upregulation of NADPH subunit Nox2 (FC vs. NG: 2.5, p<0.05). Interestingly, GCN5 inhibition or siRNA prevented hyperglycemia-induced Nox2 overexpression and subsequent O2- increase. ChIP assay showed that GCN5 binds human Nox2 promoter and mediates lysine 14 acetylation of histone 3, leading to an open chromatin and active transcription. GCN5 expression was increased in PBM of T2DM patients (FC vs. NG: 3.04, p<0.05) and significantly correlated with Nox2 transcript (r=0.32, p<0.05). Consistently, GCN5 upregulation paralleled endothelial dysfunction (r=-0.33, p<0.05) and oxidative stress (r=0.46, p<0.05), as assessed by FMD and 8-isoPGF2α, respectively. Conclusions: Epigenetic changes operated by GCN5 cause endothelial upregulation of Nox2 and subsequent oxidative stress in diabetes. Our findings provide a novel therapeutic opportunity for the prevention of diabetes-related vascular oxidative stress

Epigenetic signatures of p66Shc promoter contribute to persistent endothelial dysfunction in type 2 diabetics with optimal glycemic control

Introduction: optimal glycemic control (OGC) has failed to improve cardiovascular outcomes in patients with diabetes. We have recently reported that epigenetic regulation of the mitochondrial adaptor p66Shc, critically involved in oxidative stress, accounts for persistent endothelial dysfunction in diabetic mice with OGC. Hypothesis: we investigate whether epigenetic changes of p66Shc contribute to residual oxidative stress and vascular disease in patients with type 2 diabetes (T2DM). Methods: 7 patients with newly-diagnosed T2DM and 7 age-matched controls were studied (age 46±3 vs. 42±7 years, p=NS). After the enrolment, T2DM patients were assigned to OGC for 6 months. Glycated haemoglobin (HbA1c) and continuous blood glucose monitoring (CGM) were used as markers of glycemic control. At baseline and follow-up, patients underwent flow-mediated vasodilation (FMD) of the brachial artery. Urinary levels of 8-isoprostaglandinF2α (8-isoPGF2α) were measured as a marker of oxidative stress. p66Shc mRNA expression was assessed from peripheral blood monocytes (PBM). Chromatin immunoprecipitation (ChIP) and real time PCR were performed to investigate histone acetylation and DNA methylation of p66Shc promoter, respectively. Results: Hb1Ac (9.4±2 vs. 6.9±1%, p<0.01) and CGM (235±25 vs. 131±28 mg/dl, p<0.05) confirmed OGC in T2DM patients. As compared with controls. T2DM patients showed blunted FMD (6.8±1.9 vs. 8.6±1.4 %, p<0.05), increased urinary 8-isoPGF2α levels (295±100 vs. 33±9 pg/mL, p<0.05) and p66Shc gene upregulation (0.18±0.06 vs.0.05±0.03 AU, p<0.05). However, OGC did not rescue endothelial function (FMD 6.9±1.7 vs. 6.8±1.9%, p=NS), oxidative stress (295±100 vs. 292±85 pg/ml, p=NS) and p66Shc upregulation (0.18±0.06 vs.0.22±0.1, AU, p=NS). T2DM patients showed lysine 14 acetylation of histone 3 binding p66Shc promoter and hypomethylation of CpG dinucleotides, two critical epigenetic markers favouring p66Shc overexpression. Interestingly, these epigenetic changes remained despite OGC and correlated with persistent endothelial dysfunction and oxidative stress. Conclusions: Epigenetic regulation of p66Shc gene may contribute to the residual burden of vascular disease in T2DM individuals with OGC