In recent nanobiotechnology developments, a wide variety of functional nanomaterials and engineered biomolecules have been created, and these have numerous applications in cell biology. For these ...nanomaterials to fulfill their promises completely, they must be able to reach their biological targets at the subcellular level and with a high level of specificity. Traditionally, either nanocarrier- or membrane disruption-based method has been used to deliver nanomaterials inside cells; however, these methods are suboptimal due to their toxicity, inconsistent delivery, and low throughput, and they are also labor intensive and time-consuming, highlighting the need for development of a next-generation, intracellular delivery system. This study reports on the development of an intracellular nanomaterial delivery platform, based on unexpected cell-deformation phenomena via spiral vortex and vortex breakdown exerted in the cross- and T-junctions at moderate Reynolds numbers. These vortex-induced cell deformation and sequential restoration processes open cell membranes transiently, allowing effective and robust intracellular delivery of nanomaterials in a single step without the aid of carriers or external apparatus. By using the platform described here (termed spiral hydroporator), we demonstrate the delivery of different nanomaterials, including gold nanoparticles (200 nm diameter), functional mesoporous silica nanoparticles (150 nm diameter), dextran (hydrodynamic diameters between 2–55 nm), and mRNA, into different cell types. We demonstrate here that the system is highly efficient (up to 96.5%) with high throughput (up to 1 × 106 cells/min) and rapid delivery (∼1 min) while maintaining high levels of cell viability (up to 94%).
The regulatory mechanism of DBT-induced EPO gene expression via HIF-1α in HEK293T cell was revealed. This regulation involves two distinct mechanisms: (i) transcriptional level by increasing the ...HIF-1α mRNA and (ii) translational level by enhancing the HIF-1α protein.
Danggui Buxue Tang (DBT), a Chinese medicinal decoction that is being commonly used as hematopoietic medicine to treating woman menopausal irregularity, contains two herbs: Radix Astragali and Radix Angelicae Sinensis. Pharmacological results indicate that DBT can stimulate the production of erythropoietin (EPO), a specific hematopoietic growth factor, in cultured cells.
In order to revealing the mechanism of DBT's hematopoietic function, this study investigated the activity of the DBT-induced EPO expression and the upstream regulatory cascade of EPO via hypoxia-induced signaling in cultured kidney fibroblasts (HEK293T).
DBT-induced mRNA expressions were revealed by real-time PCR, while the change of protein expressions were analyzed by Western blotting. For the analysis of hypoxia-dependent signaling, a luciferase reporter was used to report the transcriptional activity of hypoxia response element (HRE).
The plasmid containing HRE, being transfected into HEK293T, was highly responsive to the challenge of DBT application. To account for the transcriptional activation of HRE, DBT treatment was shown to increase the mRNA and protein expressions of
hypoxia-
inducible
factor-1α (HIF-1α). In addition, the activation of Raf/MEK/ERK signaling pathway by DBT could also enhance the translation of HIF-1α, suggesting the dual actions of DBT in stimulating the EPO expression in kidney cells.
Our study indicates that HIF pathway plays an essential role in directing DBT-induced EPO expression in kidney. These results provide one of the molecular mechanisms of this ancient herbal decoction for its hematopoietic function.
Gene expression profiling (GEP) has stratified diffuse large B-cell lymphoma (DLBCL) into molecular subgroups that correspond to different stages of lymphocyte development-namely germinal center ...B-cell like and activated B-cell like. This classification has prognostic significance, but GEP is expensive and not readily applicable into daily practice, which has lead to immunohistochemical algorithms proposed as a surrogate for GEP analysis. We assembled tissue microarrays from 475 de novo DLBCL patients who were treated with rituximab-CHOP chemotherapy. All cases were successfully profiled by GEP on formalin-fixed, paraffin-embedded tissue samples. Sections were stained with antibodies reactive with CD10, GCET1, FOXP1, MUM1 and BCL6 and cases were classified following a rationale of sequential steps of differentiation of B cells. Cutoffs for each marker were obtained using receiver-operating characteristic curves, obviating the need for any arbitrary method. An algorithm based on the expression of CD10, FOXP1 and BCL6 was developed that had a simpler structure than other recently proposed algorithms and 92.6% concordance with GEP. In multivariate analysis, both the International Prognostic Index and our proposed algorithm were significant independent predictors of progression-free and overall survival. In conclusion, this algorithm effectively predicts prognosis of DLBCL patients matching GEP subgroups in the era of rituximab therapy.
Topological Dirac and Weyl semimetals not only host quasiparticles analogous to the elementary fermionic particles in high-energy physics, but also have a non-trivial band topology manifested by ...gapless surface states, which induce exotic surface Fermi arcs1,2. Recent advances suggest new types of topological semimetal, in which spatial symmetries protect gapless electronic excitations without high-energy analogues3–11. Here, using angle-resolved photoemission spectroscopy, we observe triply degenerate nodal points near the Fermi level of tungsten carbide with space group \P\bar{6}m2\ (no. 187), in which the low-energy quasiparticles are described as three-component fermions distinct from Dirac and Weyl fermions. We further observe topological surface states, whose constant-energy contours constitute pairs of ‘Fermi arcs’ connecting to the surface projections of the triply degenerate nodal points, proving the non-trivial topology of the newly identified semimetal state.
We determine and interpret fine particulate matter (PM2.5) concentrations in eastern China for January to December 2013 at a horizontal resolution of 6 km from aerosol optical depth (AOD) retrieved ...from the Korean geostationary ocean color imager (GOCI) satellite instrument. We implement a set of filters to minimize cloud contamination in GOCI AOD. Evaluation of filtered GOCI AOD with AOD from the Aerosol Robotic Network (AERONET) indicates significant agreement with mean fractional bias (MFB) in Beijing of 6.7 % and northern Taiwan of -1.2 %. We use a global chemical transport model (GEOS-Chem) to relate the total column AOD to the near-surface PM2.5. The simulated PM2.5 / AOD ratio exhibits high consistency with ground-based measurements in Taiwan (MFB =-0.52 %) and Beijing (MFB =-8.0 %). We evaluate the satellite-derived PM2.5 versus the ground-level PM2.5 in 2013 measured by the China Environmental Monitoring Center. Significant agreement is found between GOCI-derived PM2.5 and in situ observations in both annual averages (r2=0.66, N=494) and monthly averages (relative RMSE =18.3 %), indicating GOCI provides valuable data for air quality studies in Northeast Asia. The GEOS-Chem simulated chemical composition of GOCI-derived PM2.5 reveals that secondary inorganics (SO42-, NO3-, NH4+) and organic matter are the most significant components. Biofuel emissions in northern China for heating increase the concentration of organic matter in winter. The population-weighted GOCI-derived PM2.5 over eastern China for 2013 is 53.8 µg m-3, with 400 million residents in regions that exceed the Interim Target-1 of the World Health Organization.
Cognitive impairment (CI) shares common cardiovascular risk factors with acute myocardial infarction (AMI), and is increasingly prevalent in our ageing population. Whilst AMI is associated with ...increased rates of CI, CI remains underreported and infrequently identified in patients with AMI. In this review, we discuss the evidence surrounding AMI and its links to dementia and CI, including pathophysiology, risk factors, management and interventions. Vascular dysregulation plays a major role in CI, with atherosclerosis, platelet activation, microinfarcts and perivascular inflammation resulting in neurovascular unit dysfunction, disordered homeostasis and a dysfunctional neurohormonal response. This subsequently affects perfusion pressure, resulting in enlarged periventricular spaces and hippocampal sclerosis. The increased platelet activation seen in coronary artery disease (CAD) can also result in inflammation and amyloid-β protein deposition which is associated with Alzheimer’s Dementia. Post-AMI, reduced blood pressure and reduced left ventricular ejection fraction can cause chronic cerebral hypoperfusion, cerebral infarction and failure of normal circulatory autoregulatory mechanisms. Patients who undergo coronary revascularization (percutaneous coronary intervention or bypass surgery) are at increased risk for post-procedure cognitive impairment, though whether this is related to the intervention itself or underlying cardiovascular risk factors is debated. Mortality rates are higher in dementia patients with AMI, and post-AMI CI is more prevalent in the elderly and in patients with post-AMI heart failure. Medical management (antiplatelet, statin, renin-angiotensin system inhibitors, cardiac rehabilitation) can reduce the risk of post-AMI CI; however, beta-blockers may be associated with functional decline in patients with existing CI. The early identification of those with dementia or CI who present with AMI is important, as subsequent tailoring of management strategies can potentially improve outcomes as well as guide prognosis.
The KATRIN experiment aims to measure the effective electron antineutrino mass \(m_{\overline{\nu}_e}\) with a sensitivity of 0.2 eV/c\(^2\) using a gaseous tritium source combined with the MAC-E ...filter technique. A low background rate is crucial to achieving the proposed sensitivity, and dedicated measurements have been performed to study possible sources of background electrons. In this work, we test the hypothesis that gamma radiation from external radioactive sources significantly increases the rate of background events created in the main spectrometer (MS) and observed in the focal-plane detector. Using detailed simulations of the gamma flux in the experimental hall, combined with a series of experimental tests that artificially increased or decreased the local gamma flux to the MS, we set an upper limit of 0.006 count/s (90% C.L.) from this mechanism. Our results indicate the effectiveness of the electrostatic and magnetic shielding used to block secondary electrons emitted from the inner surface of the MS.
In the present studies the chronic effects of glucocorticoids and drugs activating cAMP-dependent pathways on the production and secretion of immunoreactive (ir) ANP from long term monolayer cultures ...of neonatal rat hypothalamic neurons were examined. Forskolin treatment increased ir-ANP release in a time-dependent and dose-related manner, with an EC50 of approximately 30 microM; at a lower dose of 10 microM, forskolin doubled ir-ANP release (P less than 0.01) compared to that in control cultures (mean +/- SEM, 9.6 +/- 0.3 pg/well; n = 4). While dexamethasone (DM) alone did not affect basal secretion of ir-ANP, 10 nM of the glucocorticoid significantly enhanced the effect of forskolin (10 microM) by raising ir-ANP release approximately 3 times that induced by forskolin alone (P less than 0.001). This potentiation of DM was both time dependent and dose responsive, with an EC50 of 1 nM; this effect was significantly suppressed by 100 nM RU38486, a glucocorticoid or type II receptor antagonist, but not by RU28318, a mineralocorticoid receptor antagonist. In addition, forskolin (10 microM) or DM (10 nM) alone significantly increased ir-ANP production approximately 1.4 times (P less than 0.05) and 1.3 times (P less than 0.05) over that of control cultures, respectively, whereas concurrent treatment with forskolin and DM increased ir-ANP production by approximately 1.8 times (P less than 0.01). These changes were reflected by a corresponding increment in the abundance of pro-ANP mRNA in the cultures, as demonstrated by Northern blot analysis. We conclude from the present findings that glucocorticoid- and cAMP-dependent pathways may modulate the function of ANP neurons in rat hypothalami by regulating the secretion and production of the neuropeptide at the genomic level.
We introduce a decomposition method for factorizing changes in energy demand or gas emissions over time. This method has the advantage of giving perfect decomposition. It can also handle cases with ...zero values in the data set. We compare this new method with three existing methods and summarize the respective decomposition formulae for various applications. Three application studies using data for Singapore, China, and Korea are presented. In each case, the change of a different energy or environmental indicator is decomposed using the four methods and the results obtained are compared. Our new method is superior to any of the three existing methods and may be generally applied in energy and environmental decomposition studies.
An adaptive speed controller is proposed for a permanent magnet synchronous motor (PMSM). The proposed adaptive speed regulator is insensitive to model parameter and load torque variations because it ...does not need any accurate knowledge about the motor parameter and load torque values. The stability of the proposed control system is also proven. Simulation and experimental results are presented to verify the effectiveness of the proposed adaptive speed controller under the uncertainties such as motor parameter and load torque variations using a prototype PMSM drive system.
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