Immune cells have been implicated in influencing stroke outcomes depending on their temporal dynamics, number, and spatial distribution after ischemia.Depending on their activation status, immune cells can have detrimental and beneficial properties on tissue outcome after stroke, highlighting the need to modulate inflammation towards beneficial and restorative immune responses.Novel dietary therapies may promote modulation of pro-and anti-inflammatory immune cell functions.Among the dietary interventions inspired by the Mediterranean diet, hydroxytyrosol (HT), the main phenolic component of the extra virgin olive oil (EVOO), has been suggested to have antioxidant and anti-inflammatory properties in vitro.However, immunomodulatory effects of HT have not yet been studied in vivo after stroke.The aim of this project is therefore to monitor the therapeutic effect of a HT-enriched diet in an experimental stroke model using non-invasive in vivo multimodal imaging, behavioural phenotyping and cross-correlation with ex vivo parameters.Methods: A total of N = 22 male C57BL/6 mice were fed with either a standard chow (n = 11) or a HT enriched diet (n = 11) for 35 days, following a 30 min transient middle cerebral artery occlusion (tMCAo).T2-weighted (lesion) and perfusion (cerebral blood flow)-/diffusion (cellular density)-weighted MR images were acquired at days 1, 3, 7, 14, 21 and 30 post ischemia.[ 18 F]DPA-714 (TSPO, neuroinflammation marker) PET-CT scans were acquired at days 7, 14, 21 and 30 post ischemia.Infarct volume (mm 3 ), cerebral blood flow (mL/100g/min), apparent diffusion coefficient (10 -4 •mm 2 /s) and percentage of injected tracer dose (%ID/mL) were assessed.Behavioural tests (grip test, rotarod, open field, pole test) were performed prior and after ischemia to access therapy effects on sensorimotor functions. Ivyspring International
Objective: In chronic heart failure (CHF) the myocardial expression of the inflammatory cytokine tumor necrosis factor alpha (TNF-α), which is thought to contribute to myocardial remodeling, was found to be increased. However, it is unknown whether the E3-ubiquitin ligases MAFbx and Murf-1 are involved in this remodeling process and whether their expression is regulated by TNF-α. Methods: Rats underwent ligation of the left coronary artery to induce CHF or were sham-operated. The expression of MAFbx/Murf-1 and troponin I was analyzed by RT-PCR and Western blotting in the non-infarcted area of the left ventricle. In cell culture experiments the potency of TNF-α to stimulate Murf-1/MAFbx expression, the intracellular signaling pathway, and the involvement of the E3-ligases for the impairment of contractility were assessed. Results: In CHF the myocardial expression of TNF-α was elevated 3.1-fold as compared to control. This was associated with a 4.5-fold and 2.7-fold increase in MAFbx and Murf-1 expression, respectively. A positive correlation between TNF-α and the expression of MAFbx or Murf-1 was evident. In neonatal rat cardiomyocytes, TNF-α induced the expression of MAFbx through p38MAPK-dependent pathways, whereas the induction of Murf-1 required the activation of the p42/44 MAPK pathway. Exposure of cardiomyocytes to TNF-α resulted in troponin I ubiquitinylation, subsequent degradation, and a decline in contractility. This was completely abrogated by siRNAs against Murf-1/MAFbx. Conclusion: TNF-α, which is increasingly expressed in CHF, induces troponin I degradation through a MAFbx/Murf-1-dependent pathway. This was associated with an impairment of contractility and might be one mechanism involved in the adverse remodeling process in CHF.
Human induced pluripotent stem cell (iPSC) lines have been derived from four male patients with childhood attention-deficit hyperactivity disorder (ADHD). Children and adolescents between the ages 6 and 18 suffering from ADHD were recruited for this work. Isolated keratinocytes or peripheral blood mononuclear cells from the participants were reprogrammed into iPSCs using non-integrating Sendai virus to deliver the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc.
Noninvasive monitoring of tumor therapy response helps in developing personalized treatment strategies. Here, we performed sequential PET and diffusion-weighted MRI to evaluate changes induced by a FOLFOX-like combination chemotherapy in colorectal cancer xenografts, to identify the cellular and molecular determinants of these imaging biomarkers. Methods: Tumor-bearing CD1 nude mice, engrafted with FOLFOX-sensitive Colo205 colorectal cancer xenografts, were treated with FOLFOX (5-fluorouracil, leucovorin, and oxaliplatin) weekly. On days 1, 2, 6, 9, and 13 of therapy, tumors were assessed by in vivo imaging and ex vivo analyses. In addition, HCT116 xenografts, which did not respond to the FOLFOX treatment, were imaged on day 1 of therapy. Results: In Colo205 xenografts, FOLFOX induced a profound increase in uptake of the proliferation PET tracer 3′-deoxy-3′-18F-fluorothymidine (18F-FLT) accompanied by increases in markers for proliferation (Ki-67, thymidine kinase 1) and for activated DNA damage response (γH2AX), whereas the effect on cell death was minimal. Because tracer uptake was unaltered in the HCT116 model, these changes appear to be specific for tumor response. Conclusion: We demonstrated that 18F-FLT PET can noninvasively monitor cancer treatment–induced molecular alterations, including thymidine metabolism and DNA damage response. The cellular or imaging changes may not, however, be directly related to therapy response as assessed by volumetric measurements.
Objectives To study celiac-specific antibody status over 3 years in patients with type 1 diabetes and biopsy-proven celiac disease (T1D + CD). Furthermore, to determine clinical differences after diagnosis between patients reaching constant antibody-negativity (Ab-neg) and staying antibody-positive (Ab-pos). Methods A total of 608 pediatric T1D + CD patients from the multicenter DPV registry were studied longitudinally regarding their CD specific antibody-status. Differences between Ab-neg (n = 218) and Ab-pos (n = 158) patients 3 years after biopsy were assessed and compared with 26 833 T1D patients without CD by linear and logistic regression adjusted for age, gender, diabetes duration and migration background. Results Thirty-six percent of T1D + CD patients reached and sustained antibody-negativity 3 years after CD diagnosis. The median time until patients returned to Ab-neg was 0.86 (0.51;1.16) years. Three years after diagnosis, HbA1c was lowest in Ab-neg and highest in Ab-pos patients compared to T1D-only patients (adjusted mean (95%CI): 7.72 (7.51-7.92) % vs 8.44 (8.20-8.68) % vs 8.19 (8.17-8.21) %, adjusted P < 0.001, respectively). Total cholesterol, LDL-cholesterol and frequency of dyslipidemia were significantly lower in Ab-neg compared to T1D-only patients (167 (161-173) mg/dl vs 179 (178-179) mg/dl, P < .001; 90 (84-96) mg/dl vs 99 (98-99) mg/dl, P = .005; 15.7 (10.5-22.9) % vs 25.9 (25.2-26.6) %, P = .017). In longitudinal analyses over 6 years after diagnosis, a constantly higher HbA1c (P < .001) and a lower height-SDS (P = .044) was observed in Ab-pos compared to Ab-neg patients. Conclusion Only one third of T1D + CD patients reached constant Ab-negativity after CD diagnosis. Achieving Ab-negativity after diagnosis seems to be associated with better metabolic control and growth, supposedly due to a higher adherence to therapy in general.
In vivo positron emission tomography (PET) and magnetic resonance imaging (MRI) support non-invasive assessment of the spatiotemporal expression of proteins of interest and functional/structural changes. Our work promotes the use of a volumetric analysis on multimodal imaging datasets to assess the spatio-temporal dynamics and interaction of two imaging biomarkers, with a special focus on two neuroinflammation-related biomarkers, the translocator protein (TSPO) and matrix metalloproteinases (MMPs), in the acute and chronic post-ischemic phase. To improve our understating of the neuroinflammatory reaction and tissue heterogeneity during the post ischemic phase, we aimed (i) to assess the spatio-temporal distribution of two radiotracers, [18F]DPA-714 (TSPO) and [18F]BR-351 (MMPs), (ii) to investigate their spatial interaction, including exclusive and overlapping areas, and (iii) their relationship with the T2w-MRI ischemic lesion in a transient middle cerebral artery occlusion (tMCAo) mouse model using an atlas-based volumetric analysis. As described by Zinnhardt et al. (2015), a total of N = 30 C57BL/6 mice underwent [18F]DPA-714 and [18F]BR-351 PET-CT and subsequent MR imaging 24–48 h (n = 8), 7 ± 1 days (n = 8), 14 ± 1 days (n = 7), and 21 ± 1 days (n = 7) after 30 min transient middle cerebral artery occlusion (tMCAo). To further investigate the spatio-temporal distribution of [18F]DPA-714 and [18F]BR-351, an atlas-based ipsilesional volume of interest (VOI) was applied to co-registered PET-CT images and thresholded by the mean uptake + 2.5*standard deviation of a contralateral striatal control VOI. Mean lesion-to-contralateral ratios (L/C), volume extension (V in voxel), percentages of overlap and exclusive tracer uptake areas were determined. Both tracer volumes were also compared to the lesion extent depicted by T2w-MR imaging. Both imaging biomarkers showed a constant small percentage of overlap across all time points (14.0 ± 14.2%). [18F]DPA-714 reached its maximum extent and uptake at day 14 post ischemia (V = 12,143 ± 6262 voxels, L/C = 2.32 ± 0.48). The majority of [18F]DPA-714 volume (82.4 ± 16.1%) was exclusive for [18F]DPA-714 and showed limited overlap with [18F]BR-351 and T2w-MRI lesion volumes. On the other hand, [18F]BR-351 reached its maximum extent already 24–48 h after tMCAo (V = 7279 ± 4518 voxels) and significantly decreased at day 14 (V = 1706 ± 1202 voxels). Focal spots of residual activity were still observed at day 21 post ischemia (L/C = 2.10 ± 0.37). The majority of [18F]BR-351 volume was exclusive for [18F]BR-351 (81.50 ± 25.07%) at 24–48 h and showed 64.84 ± 28.29% of overlap with [18F]DPA-714 from day 14 post ischemia while only 9.28 ± 13.45% of the [18F]BR-351 volume were overlapping the T2w-MRI lesion. The percentage of exclusive area of [18F]DPA-714 and [18F]BR-351 uptakes regarding T2w-MR lesion increased over time, suggesting that TSPO and MMPs are mostly localized in the peri‑infarct region at latter time points. This study promotes the use of an unbiased volumetric analyses of multi-modal imaging data sets to improve the characterization of pathological tissue heterogeneity. This approach improves our understanding of (i) the dynamics of disease-related multi-modal imaging biomarkers, (ii) their spatiotemporal interactions and (iii) the post-ischemic tissue heterogeneity. Our results indicate acute MMPs activation after tMCAo preceding TSPO-dependent (micro-)gliosis. The spatial distribution of MMPs and gliosis is regionally independent with only minor (< 20%) overlapping areas in peri‑infarct regions.
Myocardial perfusion imaging with single photon emission computed tomography (SPECT) is an established method for the detection and evaluation of coronary artery disease (CAD). State-of-the-art SPECT scanners yield a large number of regional parameters of the left-ventricular myocardium (e.g., blood supply at rest and during stress, wall thickness, and wall thickening during heart contraction) that all need to be assessed by the physician. Today, the individual parameters of this multivariate data set are displayed as stacks of 2D slices, bull's eye plots, or, more recently, surfaces in 3D, which depict the left-ventricular wall. In all these visualizations, the data sets are displayed side-by-side rather than in an integrated manner, such that the multivariate data have to be examined sequentially and need to be fused mentally. This is time consuming and error-prone. In this paper we present an interactive 3D glyph visualization, which enables an effective integrated visualization of the multivariate data. Results from semiotic theory are used to optimize the mapping of different variables to glyph properties. This facilitates an improved perception of important information and thus an accelerated diagnosis. The 3D glyphs are linked to the established 2D views, which permit a more detailed inspection, and to relevant meta-information such as known stenoses of coronary vessels supplying the myocardial region. Our method has demonstrated its potential for clinical routine use in real application scenarios assessed by nuclear physicians.
As a neurodevelopmental multifactorial disorder whose prevalence has been increasing worldwide, attention-deficit hyperactivity disorder (ADHD) is considered a public health concern. Methylphenidate (MPH) is the drug of choice for ADHD; however, not all patients respond fully to this treatment. Therefore, exploring the underlying molecular mechanisms involved in ADHD and potential novel therapeutic targets is crucial. Here, we generated induced pluripotent stem cells (iPSCs) from Peripheral Blood Mononuclear Cells (PBMCs) retrieved from four ADHD patients (two MPH responders and two non-responders) using Sendai virus. These lines might be helpful for the in vitro investigation of ADHD pathophysiology in a patient-specific manner.
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