46 research outputs found

    Altered adrenergic response in myocytes bordering a chronic myocardial infarction underlies <i>in vivo</i> triggered activity and repolarization instability

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    Ventricular arrhythmias are a major complication early after myocardial infarction (MI). The heterogeneous periā€infarct zone forms a substrate for reā€entry while arrhythmia initiation is often associated with sympathetic activation. We studied the mechanisms triggering these postā€MI arrhythmias in vivo and their relation to regional myocyte remodelling. In pigs with chronic MI (6 weeks), in vivo monophasic action potentials were simultaneously recorded in the periā€infarct and remote regions during adrenergic stimulation with isoproterenol (ISO). Sham animals served as controls. During infusion of ISO in vivo, the incidence of delayed afterdepolarizations (DADs) and beatā€toā€beat variability of repolarization (BVR) was higher in the periā€infarct than in the remote region. Myocytes isolated from the periā€infarct region, in comparison to myocytes from the remote region, had more DADs, associated with spontaneous Ca2+ release, and a higher incidence of spontaneous action potentials when exposed to ISO (9.99 Ā± 4.2 vs. 0.16 Ā± 0.05 APs/min, p = 0.004); these were suppressed by CaMKII inhibition. Periā€infarct myocytes also had reduced repolarization reserve and increased BVR (26 Ā± 10 ms vs. 9 Ā± 7 ms, p 2+ handling at baseline and myocyte hypertrophy were present throughout the LV. Expression of some of the related genes was however different between the regions. In conclusion, altered myocyte adrenergic responses in the periā€infarct, but not in the remote region, provide a source of triggered activity in vivo and of repolarization instability amplifying the substrate for reā€entry. These findings stimulate further exploration of regionā€specific therapies targeting myocytes and autonomic modulation

    Linear stability analysis of a horizontal phase boundary separating two miscible liquids

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    The evolution of small disturbances to a horizontal interface separating two miscible liquids is examined. The aim is to investigate how the interfacial mass transfer affects development of the Rayleigh-Taylor instability and propagation and damping of the gravity-capillary waves. The phase-field approach is employed to model the evolution of a miscible multiphase system. Within this approach, the interface is represented as a transitional layer of small but nonzero thickness. The thermodynamics is defined by the Landau free energy function. Initially, the liquid-liquid binary system is assumed to be out of its thermodynamic equilibrium, and hence, the system undergoes a slow transition to its thermodynamic equilibrium. The linear stability of such a slowly diffusing interface with respect to normal hydro- and thermodynamic perturbations is numerically studied. As a result, we show that the eigenvalue spectra for a sharp immiscible interface can be successfully reproduced for long-wave disturbances, with wavelengths exceeding the interface thickness. We also find that thin interfaces are thermodynamically stable, while thicker interfaces, with the thicknesses exceeding an equilibrium value, are thermodynamically unstable. The thermodynamic instability can make the configuration with a heavier liquid lying underneath unstable.We also find that the interfacial mass transfer introduces additional dissipation, reducing the growth rate of the Rayleigh Taylor instability and increasing the dissipation of the gravity waves. Moreover, mutual action of diffusive and viscous effects completely suppresses development of the modes with shorter wavelengths

    Uncovering a neurological protein signature for severe COVID-19

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    Coronavirus disease of 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has sparked a global pandemic with severe complications and high morbidity rate. Neurological symptoms in COVID-19 patients, and neurological sequelae post COVID-19 recovery have been extensively reported. Yet, neurological molecular signature and signaling pathways that are affected in the central nervous system (CNS) of COVID-19 severe patients remain still unknown and need to be identified. Plasma samples from 49 severe COVID-19 patients, 50 mild COVID-19 patients, and 40 healthy controls were subjected to Olink proteomics analysis of 184 CNS-enriched proteins. By using a multi-approach bioinformatics analysis, we identified a 34-neurological protein signature for COVID-19 severity and unveiled dysregulated neurological pathways in severe cases. Here, we identified a new neurological protein signature for severe COVID-19 that was validated in different independent cohorts using blood and postmortem brain samples and shown to correlate with neurological diseases and pharmacological drugs. This protein signature could potentially aid the development of prognostic and diagnostic tools for neurological complications in post-COVID-19 convalescent patients with long term neurological sequelae

    Age, Disease Severity and Ethnicity Influence Humoral Responses in a Multi-Ethnic COVID-19 Cohort

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    The COVID-19 pandemic has affected all individuals across the globe in some way. Despite large numbers of reported seroprevalence studies, there remains a limited understanding of how the magnitude and epitope utilization of the humoral immune response to SARS-CoV-2 viral anti-gens varies within populations following natural infection. Here, we designed a quantitative, multi-epitope protein microarray comprising various nucleocapsid protein structural motifs, including two structural domains and three intrinsically disordered regions. Quantitative data from the microarray provided complete differentiation between cases and pre-pandemic controls (100% sensitivity and specificity) in a case-control cohort (n = 100). We then assessed the influence of disease severity, age, and ethnicity on the strength and breadth of the humoral response in a multi-ethnic cohort (n = 138). As expected, patients with severe disease showed significantly higher antibody titers and interestingly also had significantly broader epitope coverage. A significant increase in antibody titer and epitope coverage was observed with increasing age, in both mild and severe disease, which is promising for vaccine efficacy in older individuals. Additionally, we observed significant differences in the breadth and strength of the humoral immune response in relation to ethnicity, which may reflect differences in genetic and lifestyle factors. Furthermore, our data enabled localization of the immuno-dominant epitope to the C-terminal structural domain of the viral nucleocapsid protein in two independent cohorts. Overall, we have designed, validated, and tested an advanced serological assay that enables accurate quantitation of the humoral response post natural infection and that has revealed unexpected differences in the magnitude and epitope utilization within a population

    Confounders in the assessment of the renal effects associated with low-level urinary cadmium: an analysis in industrial workers

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    <p>Abstract</p> <p>Background</p> <p>Associations of proteinuria with low-level urinary cadmium (Cd) are currently interpreted as the sign of renal dysfunction induced by Cd. Few studies have considered the possibility that these associations might be non causal and arise from confounding by factors influencing the renal excretion of Cd and proteins.</p> <p>Methods</p> <p>We examined 184 healthy male workers (mean age, 39.5 years) from a zinc smelter (n = 132) or a blanket factory (n = 52). We measured the concentrations of Cd in blood (B-Cd) and the urinary excretion of Cd (U-Cd), retinol-binding protein (RBP), protein HC and albumin. Associations between biomarkers of metal exposure and urinary proteins were assessed by simple and multiple regression analyses.</p> <p>Results</p> <p>The medians (interquartile range) of B-Cd (Ī¼g/l) and U-Cd (Ī¼g/g creatinine) were 0.80 (0.45-1.16) and 0.70 (0.40-1.3) in smelter workers and 0.66 (0.47-0.87) and 0.55 (0.40-0.90) in blanket factory workers, respectively. Occupation had no influence on these values, which varied mainly with smoking habits. In univariate analysis, concentrations of RBP and protein HC in urine were significantly correlated with both U-Cd and B-Cd but these associations were substantially weakened by the adjustment for current smoking and the residual influence of diuresis after correction for urinary creatinine. Albumin in urine did not correlate with B-Cd but was consistently associated with U-Cd through a relationship, which was unaffected by smoking or diuresis. Further analyses showed that RBP and albumin in urine mutually distort their associations with U-Cd and that the relationship between RBP and Cd in urine was almost the replicate of that linking RBP to albumin</p> <p>Conclusions</p> <p>Associations between proteinuria and low-level urinary Cd should be interpreted with caution as they appear to be largely driven by diuresis, current smoking and probably also the co-excretion of Cd with plasma proteins.</p

    Novel engineered nanobodies specific for N-terminal region of alpha-synuclein recognize Lewy-body pathology and inhibit in-vitro seeded aggregation and toxicity.

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    Nanobodies (Nbs), the single-domain antigen-binding fragments of dromedary heavy-chain antibodies (HCAb), are excellent candidates as therapeutic and diagnostic tools in synucleinopathies because of their small size, solubility and stability. Here, we constructed an immune nanobody library specific to the monomeric form of alpha-synuclein (Ī±-syn). Phage display screening of the library allowed the identification of a nanobody, NbĪ±-syn01, specific for Ī±-syn. Unlike previously developed nanobodies, NbĪ±-syn01 recognized the N-terminal region which is critical for inā€‰vitro and inā€‰vivo aggregation and contains many point mutations involved in early PD cases. The affinity of the monovalent NbĪ±-syn01 and the engineered bivalent format BivNbĪ±-syn01 measured by isothermal titration calorimetry revealed unexpected results where NbĪ±-syn01 and its bivalent format recognized preferentially Ī±-syn fibrils compared to the monomeric form. NbĪ±-syn01 and BivNbĪ±-syn01 were also able to inhibit Ī±-syn-seeded aggregation inā€‰vitro and reduced Ī±-syn-seeded aggregation and toxicity in cells showing their potential to reduce Ī±-syn pathology. Moreover, both nanobody formats were able to recognize Lewy-body pathology in human post-mortem brain tissue from PD and DLB cases. Additionally, we present evidence through structural docking that NbĪ±-syn01 binds the N-terminal region of the Ī±-syn aggregated form. Overall, these results highlight the potential of NbĪ±-syn01 and BivNbĪ±-syn01 in developing into a diagnostic or a therapeutic tool for PD and related disorders

    Fibrillar form of Ī±-synuclein-specific scFv antibody inhibits Ī±-synuclein seeds induced aggregation and toxicity

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    Synucleinopathies including Parkinsonā€™s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA) are characterized by pathological accumulation of Ī±-synuclein (Ī±-syn). Amongst the various approaches attempting to tackle the pathological features of synucleinopathies, antibody-based immunotherapy holds much promise. However, the large size of antibodies and corresponding difficulty in crossing the blood-brainĀ barrier has limited development in this area. To overcome this issue, we engineered single-chain variable fragments (scFvs) against fibrillar Ī±-syn, a putative disease-relevant form of Ī±-syn. The purified scFvs showed specific activity towards Ī±-syn fibrils and oligomers in comparison to monomers and recognized intracellular inclusions in human post-mortem brain tissue of Lewy body disease cases, but not aged controls. In vitro studies indicated scFvs inhibit the seeding of Ī±-syn aggregation in a time-dependent manner, decreased Ī±-syn seed-induced toxicity in a cell model of PD, and reduced the production of insoluble Ī±-syn phosphorylated at Ser-129 (pS129-Ī±-syn). These results suggest that our Ī±-syn fibril-specific scFvs recognize Ī±-syn pathology and can inhibit the aggregation of Ī±-syn in vitro and prevent seeding-dependent toxicity. Therefore, the scFvs described here have considerable potential to be utilized towards immunotherapy in synucleinopathies and may also have applications in ante-mortem imaging modalities.Dr. El-Agnafā€™s laboratory was funded by Qatar Biomedical Research Institute under the Start-up Fund SF 2017ā€“ 007. The Newcastle Brain Tissue Resource is funded in part by a grant from the UK Medical Research Council, by NIHR Newcastle Biomedical Research Centre awarded to the Newcastle upon Tyne NHS Foundation Trust and Newcastle University, and by a grant from the Alzheimerā€™s Society and Alzheimerā€™s Research UK as part of the Brains for Dementia Research Project

    Signal transducer and activator of transcription 3 (Stat3) suppresses stat1/interferon signaling pathway and inflammation in senescent preadipocytes

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    Ā© 2021 by the authors. Licensee MDPI, Basel, Switzerland. Obesity promotes premature aging and dysfunction of white adipose tissue (WAT) through the accumulation of cellular senescence. The senescent cells burden in WAT has been linked to inflammation, insulinā€resistance (IR), and type 2 diabetes (T2D). There is limited knowledge about molecular mechanisms that sustain inflammation in obese states. Here, we describe a robust and physiologically relevant in vitro system to trigger senescence in mouse 3T3ā€L1 preadipocytes. By employing transcriptomics analyses, we discovered upā€regulation of key proā€inflammatory molecules and activation of interferon/signal transducer and activator of transcription (STAT)1/3 signaling in senescent preadipocytes, and expression of downstream targets was induced in epididymal WAT of obese mice, and obese human adipose tissue. To test the relevance of STAT1/3 signaling to preadipocyte senescence, we used Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) technology to delete STAT1/3 and discovered that STAT1 promoted growth arrest and cooperated with cyclic Guanosine Monophosphateā€Adenosine Monophosphate (GMPā€AMP) synthaseā€stimulator of interferon genes (cGASā€STING) to drive the expression of interferon Ī² (IFNĪ²), Cā€Xā€C motif chemokine ligand 10 (CXCL10), and interferon signalingā€related genes. In contrast, we discovered that STAT3 was a negative regulator of STAT1/cGASā€STING signalingā€”it suppressed senescence and inflammation. These data provide insights into how STAT1/STAT3 signaling coordinates senescence and inflammation through functional interactions with the cGAS/STING pathway
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