Nanomechanical properties of α-synuclein amyloid fibrils: a comparative study by nanoindentation, harmonic force microscopy, and Peakforce QNM

We report on the use of three different atomic force spectroscopy modalities to determine the nanomechanical properties of amyloid fibrils of the human α-synuclein protein. α-Synuclein forms fibrillar nanostructures of approximately 10 nm diameter and lengths ranging from 100 nm to several microns, which have been associated with Parkinson's disease. Atomic force microscopy (AFM) has been used to image the morphology of these protein fibrils deposited on a flat surface. For nanomechanical measurements, we used single-point nanoindentation, in which the AFM tip as the indenter is moved vertically to the fibril surface and back while the force is being recorded. We also used two recently developed AFM surface property mapping techniques: Harmonic force microscopy (HarmoniX) and Peakforce QNM. These modalities allow extraction of mechanical parameters of the surface with a lateral resolution and speed comparable to tapping-mode AFM imaging. Based on this phenomenological study, the elastic moduli of the α-synuclein fibrils determined using these three different modalities are within the range 1.3-2.1 GPa. We discuss the relative merits of these three methods for the determination of the elastic properties of protein fibrils, particularly considering the differences and difficulties of each method

DMA, a Bisbenzimidazole, Offers Radioprotection by Promoting NFκB Transactivation through NIK/IKK in Human Glioma Cells

BACKGROUND: Ionizing radiation (IR) exposure often occurs for human beings through occupational, medical, environmental, accidental and/or other sources. Thus, the role of radioprotector is essential to overcome the complex series of overlapping responses to radiation induced DNA damage. METHODS AND RESULTS: Treatment of human glioma U87 cells with DMA (5- {4-methylpiperazin-1-yl}-2-[2'-(3, 4-dimethoxyphenyl)-5'-benzimidazolyl] in the presence or absence of radiation uncovered differential regulation of an array of genes and proteins using microarray and 2D PAGE techniques. Pathway construction followed by relative quantitation of gene expression of the identified proteins and their interacting partners led to the identification of MAP3K14 (NFκB inducing kinase, NIK) as the candidate gene affected in response to DMA. Subsequently, over expression and knock down of NIK suggested that DMA affects NFκB inducing kinase mediated phosphorylation of IKKα and IKKβ both alone and in the presence of ionizing radiation (IR). The TNF-α induced NFκB dependent luciferase reporter assay demonstrated 1.65, 2.26 and 3.62 fold increase in NFκB activation at 10, 25 and 50 µM DMA concentrations respectively, compared to control cells. This activation was further increased by 5.8 fold in drug + radiation (50 µM +8.5 Gy) treated cells in comparison to control. We observed 51% radioprotection in control siRNA transfected cells that attenuated to 15% in siRNA NIK treated U87 cells, irradiated in presence of DMA at 24 h. CONCLUSIONS: Our studies show that NIK/IKK mediated NFκB activation is more intensified in cells over expressing NIK and treated with DMA, alone or in combination with ionizing radiation, indicating that DMA promotes NIK mediated NFκB signaling. This subsequently leads to the radioprotective effect exhibited by DMA

Sex differences in atrial fibrillation in India: Insights from the Kerala-AF registry

Background: Much data informing sex differences in atrial fibrillation (AF) comes from Western cohorts. In this analysis, we describe sex differences in Kerala, India, using the Kerala-AF registry—the largest AF registry from the Indian subcontinent. Methods: Patients aged ≥18 years were recruited from 53 hospitals across Kerala. Patients were compared for demographics, treatments, and 12-month outcomes, including major adverse cardiovascular events (MACE) and bleeding. Results: Male patients were more likely to have a smoking and/or alcohol history and had more ischaemic heart disease (46.2% vs. 25.5%; p < 0.001). Female patients had more valvular AF (35.1% vs. 18.0%; p < 0.001), and more use of calcium-channel blockers (23.3% vs. 16.5%; p < 0.001) or digoxin (39.6% vs. 28.5%; p < 0.001). Almost one in four patients were not anticoagulated despite raised CHA2DS2-VASc scores. 12-month MACE outcomes did not differ by sex (male: 30.2% vs. female: 29.4%; p = 0.685), though bleeding events were more common in male patients (2.4% vs. 1.3%; p = −0.038), driven by minor bleeding (1.2% vs. 0.5%). Conclusion: In this large AF cohort from India, male patients had a higher prevalence of ischaemic heart disease, smoking, and alcohol use, while female patients had a higher prevalence of valvular heart disease. MACE did not differ by sex, though bleeding was more common in males. Almost a quarter of patients were not anticoagulated despite raised thromboembolic risk

Phenotypes of South Asian patients with atrial fibrillation and holistic integrated care management: cluster analysis of data from KERALA-AF Registry

Background: Patients with atrial fibrillation (AF) frequently experience multimorbidity. Cluster analysis, a machine learning method for classifying patients with similar phenotypes, has not yet been used in South Asian AF patients. Methods: The Kerala Atrial Fibrillation Registry is a prospective multicentre cohort study in Kerala, India, and the largest prospective AF registry in South Asia. Hierarchical clustering was used to identify different phenotypic clusters. Outcomes were all-cause mortality, major adverse cardiovascular events (MACE), and composite bleeding events within one-year follow-up. Findings: 3348 patients were included (median age 65.0 [56.0–74.0] years; 48.8% male; median CHA2DS2-VASc 3.0 [2.0–4.0]). Five clusters were identified. Cluster 1: patients aged ≤65 years with rheumatic conditions; Cluster 2: patients aged >65 years with multi-comorbidities, suggestive of cardiovascular-kidney-metabolic syndrome; Cluster 3: patients aged ≤65 years with fewer comorbidities; Cluster 4: heart failure patients with multiple comorbidities; Cluster 5: male patients with lifestyle-related risk factors. Cluster 1, 2 and 4 had significantly higher MACE risk compared to Cluster 3 (Cluster 1: OR 1.36, 95% CI 1.08–1.71; Cluster 2: OR 1.79, 95% CI 1.42–2.25; Cluster 4: OR 1.76, 95% CI 1.31–2.36). The results for other outcomes were similar. Atrial fibrillation Better Care (ABC) pathway in the whole cohort was low (10.1%), especially in Cluster 4 (1.9%). Overall adherence to the ABC pathway was associated with reduced all-cause mortality (OR 0.26, 95% CI 0.15–0.46) and MACE (OR 0.45, 95% CI 0.31–0.46), similar trends were evident in different clusters. Interpretation: Cluster analysis identified distinct phenotypes with implications for outcomes. There was poor ABC pathway adherence overall, but adherence to such integrated care was associated with improved outcomes. Funding: Kerala Chapter of Cardiological Society of India

Effect of Inlet Tangential port area on the Performance of Small Scale Simplex Atomizer

An experimental investigation was conducted to study the effects of increased area of inlet tangential ports on the performance of small scale simplex atomizer. The spray characteristics of three different simplex atomizer representing varying area of inlet tangential ports are examined using water as a working fluid. Measurements of coefficient of discharge, spray cone angle, Sauter mean diameter and droplet size distribution were carried out over wide range of injection pressure. Coriolis mass flow meter was used to measure coefficient of discharge. Spray cone angle was measured by image processing technique. Sauter mean diameter and droplet size distributions were measured by Malvern droplet sizing instrument. It was observed that with increase in area of inlet tangential ports the size of air core produced along the center line reduced, which increases the coefficient of discharge. Spray cone angle decreases with increase in area of inlet tangential ports reduces swirl strength inside swirl chamber, which results in increasing Sauter mean diameter. Better droplet size distribution was observed for lower area of inlet tangential port configuration. Good agreement was observed between the obtained experimental results and experimental correlations available in literatures

Identification of putative QTLs for seedling stage phosphorus starvation response in finger millet (Eleusine coracana L. Gaertn.) by association mapping and cross species synteny analysis

A germplasm assembly of 128 finger millet genotypes from 18 countries was evaluated for seedling-stage phosphorus (P) responses by growing them in P sufficient (Psuf) and P deficient (Pdef) treatments. Majority of the genotypes showed adaptive responses to low P condition. Based on phenotype behaviour using the best linear unbiased predictors for each trait, genotypes were classified into, P responsive, low P tolerant and P non-responsive types. Based on the overall phenotype performance under Pdef, 10 genotypes were identified as low P tolerants. The low P tolerant genotypes were characterised by increased shoot and root length and increased root hair induction with longer root hairs under Pdef, than under Psuf. Association mapping of P response traits using mixed linear models revealed four quantitative trait loci (QTLs). Two QTLs (qLRDW.1 and qLRDW.2) for low P response affecting root dry weight explained over 10% phenotypic variation. In silico synteny analysis across grass genomes for these QTLs identified putative candidate genes such as Ser-Thr kinase and transcription factors such as WRKY and basic helix-loop-helix (bHLH). The QTLs for response under Psuf were mapped for traits such as shoot dry weight (qHSDW.1) and root length (qHRL.1). Putative associations of these QTLs over the syntenous regions on the grass genomes revealed proximity to cytochrome P450, phosphate transporter and pectin methylesterase inhibitor (PMEI) genes. This is the first report of the extent of phenotypic variability for P response in finger millet genotypes during seedling-stage, along with the QTLs and putative candidate genes associated with P starvation toleranc

A Population-based Model of Local Control and Survival Benefit of Radiotherapy for Lung Cancer

Aims To estimate the population-based locoregional control and overall survival benefits of radiotherapy for lung cancer if the whole population were treated according to evidence-based guidelines. These estimates were based on a published radiotherapy utilisation (RTU) model that has been used to estimate the demand and planning of radiotherapy services nationally and internationally. Materials and methods The lung cancer RTU model was extended to incorporate an estimate of benefits of radiotherapy alone, and of radiotherapy in conjunction with concurrent chemotherapy (CRT). Benefits were defined as the proportional gains in locoregional control and overall survival from radiotherapy over no radiotherapy for radical indications, and from postoperative radiotherapy over surgery alone for adjuvant indications. A literature review (1990–2015) was conducted to identify benefit estimates of individual radiotherapy indications and summed to estimate the population-based gains for these outcomes. Model robustness was tested through univariate and multivariate sensitivity analyses. Results If evidence-based radiotherapy recommendations are followed for the whole lung cancer population, the model estimated that radiotherapy alone would result in a gain of 8.3% (95% confidence interval 7.4–9.2%) in 5 year locoregional control, 11.4% (10.8–12.0%) in 2 year overall survival and 4.0% (3.6–4.4%) in 5 year overall survival. For the use of CRT over radiotherapy alone, estimated benefits would be: locoregional control 1.7% (0.8–2.4%), 2 year overall survival 1.7% (0.5–2.8%) and 5 year overall survival 1.2% (0.7–1.9%). Conclusions The model provided estimates of radiotherapy benefit that could be achieved if treatment guidelines are followed for all cancer patients. These can be used as a benchmark so that the effects of a shortfall in the utilisation of radiotherapy can be better understood and addressed. The model can be adapted to other populations with known epidemiological parameters to ensure the planning of equitable radiotherapy services