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Endoplasmic reticulum (ER) stress and unfolded protein response are cells' survival strategies to thwart disruption of proteostasis. Tumor cells are continuously being challenged by ER stress. The prion protein, PrP, normally a glycosylphosphatidylinositol (GPI)-anchored protein exists as a pro-PrP retaining its GPI-peptide signal sequence in human pancreatic ductal cell adenocarcinoma (PDAC). Higher abundance of pro-PrP indicates poorer prognosis in PDAC patients. The reason why PDAC cells express pro-PrP is unknown. Here, we report that persistent ER stress causes conversion of GPI-anchored PrP to pro-PrP via a conserved ATF6-miRNA449c-5p-PIGV axis. Mouse neurons and AsPC-1, a PDAC cell line, express GPI-anchored PrP. However, continuous culture of these cells with the ER stress inducers thapsigargin or brefeldin A results in the conversion of a GPI-anchored PrP to pro-PrP. Such a conversion is reversible; removal of the inducers allows the cells to re-express a GPI-anchored PrP. Mechanistically, persistent ER stress increases the abundance of an active ATF6, which increases the level of miRNA449c-5p (miR449c-5p). By binding the mRNA of PIGV at its 3'-UTRs, miR449c-5p suppresses the level of PIGV, a mannosyltransferase pivotal in the synthesis of the GPI anchor. Reduction of PIGV leads to disruption of the GPI anchor assembly, causing pro-PrP accumulation and enhancing cancer cell migration and invasion. The importance of ATF6-miR449c-5p-PIGV axis is recapitulated in PDAC biopsies as the higher levels of ATF6 and miR449c-5p and lower levels of PIGV are markers of poorer outcome for patients with PDAC. Drugs targeting this axis may prevent PDAC progression.
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Adenocarcinoma , Carcinoma Ductal Pancreático , Estresse do Retículo Endoplasmático , Glicosilfosfatidilinositóis , Neoplasias Pancreáticas , Proteínas Priônicas , Animais , Humanos , Camundongos , Fator 6 Ativador da Transcrição/genética , Adenocarcinoma/patologia , Glicosilfosfatidilinositóis/metabolismo , Neoplasias Pancreáticas/metabolismo , Proteínas Priônicas/genética , Proteínas Priônicas/metabolismo , Neoplasias PancreáticasRESUMO
Metasurface zone plates exhibit stronger optical control capabilities than traditional Fresnel zone plates, especially in polarization transformation and multiplexing. However, there are still few studies on metasurface zone plates that can be used for simultaneous control of forward and backward waves. In this work, we propose what is to our knowledge a new scheme that utilizes metasurface zone plates for orthogonal linear polarization separation and wavefront manipulation at the same time. We demonstrate the separation of linearly polarized components and transmission-reflection focusing by using the destructive and constructive interference between different meta-atoms in the super-cell, as well as the phase difference between the super-cells. The metasurface not only needs a simple binary phase design but also shows a working bandwidth more than 30â nm with a central wavelength of 875â nm. This scheme can be extended to other electromagnetic bands such as visible and terahertz ones, providing an important way for the multi-dimensional light field manipulations.
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Circular dichroism (CD) spectrum and optical rotation (OR) spectrum, crucial for understanding molecular properties and configurations, present challenges due to limited testing methods and equipment accuracy in the ultraviolet (UV) region. This study proposes a weak measurement system for chiral signals in varying concentrations in the ultraviolet range, optimized using a deep neural network (DNN) model. Introducing different post-selections to detect the circular dichroism spectrum and optical rotation spectrum separately, with contrast as a probe, it achieves a detection resolution of up to 10-6 rad. Moreover, the fitted value of the training data can reach 0.9989, enhancing the prediction accuracy of chiral molecule concentrations. This method exhibits considerable promise for applications in chiral measurement and sensor technologies.
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With the rapid development of 5G communication technology, microwave dielectric ceramics with high dielectric constants are very conducive to the miniaturization of passive devices. Here, Ba3Ti4-x(Ni1/3Nb2/3)xNb4O21 (BTN â¼ NN, 0.03 ≤ x ≤ 0.15) ceramics with hexagonal phases are synthesized via the solid-phase route. The composite (Ni1/3Nb2/3)4+ ion substitution strategy can substantially improve the microwave dielectric properties of the Ba3Ti4Nb4O21 (BTN) ceramic. The εr and Q × f values depend on the ionicity (Nb-O bonds) and lattice energies (Nb(1)-O3 and Nb(1)-O2(1) bonds). The microwave dielectric properties of the BTN â¼ NN (x = 0.09) ceramic sintered at 1250 °C are εr = 60.3, Q × f = 22073 GHz, and τf = 78.1 ppm/°C. A miniaturized all-ceramic radome (@400 mm × 400 mm × 8 mm) for 5G beam-splitting function is designed and demonstrated using this ceramic. Compared to other radomes designed for other work utilizing low εr, the size of this radome has been reduced by 3/7. The reflection coefficients of the beam splitting function are all 0.73, and the phase shifts are all 360°. This work contributes to the development of miniaturized passive devices from a materials point of view.
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We design a broadband free space 2×4 90° optical hybrid over a spectral window of 1000-1200 nm and 1470-1650 nm and verify the feasibility of the scheme experimentally. The hybrid consists of three broadband polarization beam splitters, an achromatic λ/4 wave plate, and three achromatic λ/2 wave plates. The fabricated hybrid exhibits a good quadrature phase response with an interchannel imbalance of 0.93-1.07 and a low phase deviation of less than 0.2° under the typical communication wavelengths of 1064 nm and C-band. The experimental results of the heterodyne method show that the proposed hybrid can effectively solve the wavelength incompatibility problem in satellite laser communication and realize interconnection at different wavelengths. The designed hybrid (without coupling) has a measured insertion loss of no more than 7.34 dB at 1064 nm and C-band. A high-speed transmission experiment with BPSK format has been conducted to verify the performance of the assembled device.
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There are three isoforms of human collagen prolyl 4-hydroxylases (C-P4Hs), each of which has been reported to play an important role in regulating the progression of a variety of human cancers. By analyzing TGCA datasets on human head and neck squamous cell carcinoma (HNSC), we find that a higher expression of all three C-P4HAs (the α subunit of C-P4Hs) is a superior prognostic indicator than a higher expression of two or a single C-P4HA. Unexpectedly, some patients with higher levels of three C-P4HAs survive longer than patients whose tumors have lower expression of C-P4HAs. Therefore, there may be molecule(s) that can negate the deleterious effects of overexpressing C-P4HAs during cancer progression. By constructing a functional protein interaction network of C-P4HAs and analyzing molecules whose expressions are correlated significantly with that of C-P4HAs, we identify scribble cell polarity complex component 2 (LLGL2) as a factor that antagonizes the effects of overexpressed C-P4HAs on HNSC. Silencing of LLGL2 in the human oral squamous cell line Cal-27 upregulates the expression of occludin and increases cancer cell invasion and migration. In contrast, knocking down C-P4HA alone inhibits cell migration and invasion. Furthermore, simultaneously downregulating three C-P4HAs has more pronounced effects on inhibiting cell migration and invasion. Accordingly, high LLGL2 expression is also a marker indicating improved prognosis in patients with HNSC. These results suggest that the interplay between LLGL2 and C-P4HAs may be targeted to mitigate HNSC tumorigenesis and progression.
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The Internet of Things (IoT) plays an essential role in people's daily lives, such as healthcare, home, traffic, industry, and so on. With the increase in IoT devices, there emerge many security issues of data loss, privacy leakage, and information temper in IoT network applications. Even with the development of quantum computing, most current information systems are weak to quantum attacks with traditional cryptographic algorithms. This paper first establishes a general security model for these IoT network applications, which comprises the blockchain and a post-quantum secure identity-based signature (PQ-IDS) scheme. This model divides these IoT networks into three layers: perceptual, network, and application, which can protect data security and user privacy in the whole data-sharing process. The proposed PQ-IDS scheme is based on lattice cryptography. Bimodal Gaussian distribution and the discrete Gaussian sample algorithm are applied to construct the fundamental difficulty problem of lattice assumption. This assumption can help resist the quantum attack for information exchange among IoT devices. Meanwhile, the signature mechanism with IoT devices' identity can guarantee non-repudiation of information signatures. Then, the security proof shows that the proposed PQ-IDS can obtain the security properties of unforgeability, non-repudiation, and non-transferability. The efficiency comparisons and performance evaluations show that the proposed PQ-IDS has good efficiency and practice in IoT network applications.
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AZO-coated ZnO core-shell nanorods were successfully fabricated using the mist chemical vapor deposition method. The influence of coating time on the structural, optical, and photocatalytic properties of zinc oxide nanorods was investigated. It was observed that the surface area of AZO-coated ZnO core-shell nanorods increased with an increase in coating time. The growth orientation along the (0001) crystal plane of the AZO thin film coating was the same as that of zinc oxide nanorods. The crystallinity of AZO-coated ZnO core-shell nanorods was significantly improved as well. The optical transmittance of AZO-coated ZnO core-shell nanorods was greater than 55% in the visible region. The degradation efficiency for methyl red dye solution increased with an increase in coating time. The highest degradation efficiency was achieved by AZO-coated ZnO core-shell nanorods with a coating duration of 20 min, exhibiting a degradation rate of 0.0053 min-1. The photodegradation mechanism of AZO-coated ZnO core-shell nanorods under ultraviolet irradiation was revealed.
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OBJECTIVES: To examine whether the associations of neighborhood environment and body mass index (BMI) of community-dwelling older adults aged 70 and above were mediated by transport-related physical activity (TRPA). METHODS: A bootstrap method was employed to test the mediation model with multisource data from Chongqing, China. RESULTS: Neighborhood walkability (effect: 0.030, 95% CI [0.001-0.160]) and shopping facility accessibility (effect: 0.002, 95 % CI [0.001 - 0.101]) exhibited positive effects on BMI indirectly through decreasing TRPA duration. Negative indirect effects of sports facility accessibility (effect: -0.004, 95 % CI [-0.112 - -0.003]) and transit accessibility (effect: -0.044, 95 % CI [-0.074 - -0.002]) on BMI were observed through increasing TRPA duration. Park accessibility showed both direct (effect: -0.242, p < 0.05) and indirect (effect: -0.036, 95 % CI [-0.061 - -0.005]) negative correlations with BMI. CONCLUSION: Our findings facilitate neighborhood environment interventions regarding obesity among older adults in developing countries.
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Exercício Físico , Vida Independente , Humanos , Idoso , Índice de Massa Corporal , Características de Residência , China , Características da Vizinhança , CaminhadaRESUMO
In this paper, we propose a precision method to measure the chiroptical signal of Artemisinin solutions using the photonic spin Hall effect (PSHE) on the Ce:YIG-YIG-SiO2 structure as a probe. The effects of transmission distance, incident angles, applied magnetic fields of different directions, and beam waist of light on the weak measurement system are analytically investigated through simulations. It is found that decreasing the beam waist of the incident spot, increasing the incident angle, increasing the transmission distance, and adding a longitudinal magnetic field is conducive to enhancing the amplification transverse shift of PSHE, thus the measurement sensitivity is greatly improved. Based on the optimal weak measurement scheme, the detection limit for concentration measurement of artemisinin based on optical rotatory (OR) was reduced to 0.05 mg/ml. The measurement precision of the OR angle has been improved to 10-7rad.
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Hyperbolic metamaterial (HMM) based sensors can achieve superior sensing performance than conventional surface plasmon resonance sensors. In this work, the operator approach to effective medium approximation (OEMA) is used to characterize the HMM dielectric constant properties of metal-dielectric multilayer structures, which are classified into short-range (SR) mode and long-range (LR) mode according to the propagation length of the bulk high K waves in HMM. The dispersion relations of SR modes are derived, and a high-sensitivity refractive index sensor is designed for the near-infrared SR mode resonance. The effects of the number of periods, cell thickness, metal fill rate and incidence angle on the SR mode resonance were analyzed for the multilayer structured HMM. Our designed sensing structure achieves a maximum sensitivity of 330â µm/RIU in the near-infrared band with a quality factor of 492 RIU-1. In addition, the simulations show that the SR mode resonance wavelength is flexible and tunable. We believe that the study of HMM-based SR mode resonance sensors offers potential applications for high-sensitivity biochemical detection.
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Traditional grating lenses can accumulate phase for adjusting wavefronts, and plasmonic resonances can be excited in metasurfaces with discrete structures for optical field modulation. Diffractive and plasma optics have been developing in parallel, with easy processing, small size, and dynamic control advantages. Due to theoretical hybridization, structural design can combine advantages and show great potential value. Changing the shape and size of the flat metasurface can easily produce light field reflections, but changes in height are rarely cross-explored. We propose a graded metasurface with a single-structure periodic arrangement, which can mix the effects of plasmonic resonance and grating diffraction. As for solvents of different polarities, strong polarization-dependent beam reflections are produced, enabling versatile beam convergence and deflection. Dielectric/metal nanostructures with selective hydrophobic/hydrophilic properties can be arranged by the structural material specification to selectively settle the location of the solution in a liquid environment. Furthermore, the wetted metasurface is actively triggered to achieve spectral control and initiate polarization-dependent beam steering in the broadband visible light region. Actively reconfigurable polarization-dependent beam steering has potential applications in tunable optical displays, directional emission, beam manipulation and processing, and sensing technologies.
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Retinal pigment epithelial (RPE) cellular senescence is regarded as an initiator for age-related macular degeneration (AMD). We previously demonstrated that by the coculture way, embryonic stem cells (ESCs) can reverse the senescence of RPE cells, but xenograft cells can cause a plethora of adverse effects. Extracellular vesicles (EVs) derived from ESCs can act as messengers to mediate nearby cell activities and have the same potential as ESCs to reverse RPE senescence. Furthermore, ESC-EVs have achieved preliminary efficacy while treating many age-related diseases. The present study aimed to test the effect of ESC-EVs on the replicative senescence model of RPE cells as well as its mechanism. The results showed that ESC-EVs enhanced the proliferative ability and cell cycle transition of senescent RPE cells, whereas reduced the senescence-associated galactosidase (SA-ß-gal) staining rate, as well as the levels of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). Moreover, classical markers of cellular senescence p21WAF1/CIP1 (p21) and p16INK4a (p16) were downregulated. The bioinformatic analysis and further study showed that the inhibition of the p38MAPK pathway by ESC-EVs played a pivotal role in RPE cellular senescence-reversing effect, which was ameliorated or even abolished when dehydrocorydaline were administrated simultaneously, demonstrating that ESC-EVs can effectively reverse RPE cellular senesence by inhibiting the p38MAPK pathway, thus highlights the potential of ESC-derived EVs as biomaterials for preventative and protective therapy in AMD.
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Vesículas Extracelulares , Epitélio Pigmentado da Retina , Humanos , Epitélio Pigmentado da Retina/metabolismo , Células-Tronco Embrionárias , Células Epiteliais , Pigmentos da Retina/metabolismo , Senescência CelularRESUMO
We report an efficient alkyl transfer strategy for the direct ß-alkylation of chalcones using commercially available alkyl bromides as alkyl reagents. In this transformation, the ortho-phosphanyl substituent in the chalcones is crucial for controlling their reactivity and selectivity. It also serves as a reliable alkyl transfer shuttle to transform electrophilic alkyl bromides into nucleophilic alkyl species in the form of quaternary phosphonium salts and transfer the alkyl group effectively to the ß-position of the chalcones. This alkyl transfer strategy can be further extended to the alkenylation of ortho-phosphanyl benzaldehydes to assemble functionalized polyenes.
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Chalconas , Brometos , Catálise , Sais , AlquilaçãoRESUMO
BACKGROUND: Depression affects 10%-20% of older adults worldwide. The course of late-life depression (LLD) is often chronic, with a poor long-term prognosis. Lower treatment adherence, stigma, and suicide risk lead to significant challenges in the continuity of care (COC) for patients with LLD. Elderly patients with chronic diseases can benefit from COC. As a common chronic disease of the elderly, whether depression can also benefit from COC has not been systematically reviewed. METHODS: Systematic literature search in Embase, Cochrane Library, Web of Science, Ovid, PubMed and Medline. Randomized Controlled Trials (RCTs) on the intervention effects of COC and LLD, published on 12 April 2022, were selected. Two independent researchers made research choices based on consensus. An RCT with COC as an intervention measure for the elderly with depression 60 years old was the inclusion criteria. RESULTS: A total of 10 RCTs involving 1557 participants were identified in this study. The findings showed that: (1) COC significantly reduced depressive symptoms compared to usual care (standardized mean difference [SMD] = -0.47, 95% confidence interval: -0.63 to -0.31), with the best improvement at 3- to 6-month follow-up; (2) The reduction in depressive symptoms was more pronounced for patients with comorbid chronic conditions with LLD (SMD = -0.93, 95% CI: -1.18 to -0.68); (3) COC was more effective than other regions for LLD in Europe and the Americas (SMD = -0.84, 95% CI: -1.07 to -0.61); and (4) COC had a positive impact on the quality of life of patients with LLD (SMD = 0.21, 95% CI: 0.02-0.40). LIMITATIONS: The included studies included several multi-component interventions with widely varying methods. Therefore, it was almost impossible to analyze which of these interventions had an impact on the assessed outcomes. CONCLUSIONS: This meta-analysis shows that COC can significantly reduce depressive symptoms and improve quality of life in patients with LLD. However, when treating and caring for patients with LLD, health care providers should also pay attention to timely adjustments of intervention plans according to follow-up, synergistic interventions for multiple co-morbidities, and actively learning from advanced COC programs at home and abroad to improve the quality and effectiveness of services.
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Depressão , Qualidade de Vida , Humanos , Idoso , Depressão/terapia , Comorbidade , Doença Crônica , Europa (Continente)RESUMO
BACKGROUND: This meta-analysis aimed to explore the impact of red blood cell (RBC) transfusion on mortality during extracorporeal membrane oxygenation (ECMO). Previous studies investigated the prognostic impact of RBC transfusion during ECMO on the risk of mortality, but no meta-analysis has been published before. METHODS: The PubMed, Embase, and the Cochrane library were systematically searched for papers published up to 13 December 2021, using the MeSH terms "ECMO", "'Erythrocytes", and "Mortality" to identify meta-analyses. Total or daily RBC transfusion during ECMO and mortality were examined. RESULTS: The random-effect model was used. Eight studies (794 patients, including 354 dead) were included. The total volume of RBC was associated with higher mortality standardized weighted difference (SWD = -0.62, 95% CI: -1.06,-0.18, p = .006; I2 = 79.7%, Pheterogeneity = 0.001). The daily volume of RBC was associated with higher mortality (SWD = -0.77, 95% CI: -1.11,-0.42, p < .001; I2 = 65.7%, Pheterogeneity = 0.020). The total volume of RBC was associated with mortality for venovenous (VV) (SWD = -0.72, 95% CI: -1.23, -0.20, p = .006) but not venoarterial ECMO (p = .126) or when reported together (p = .089). The daily volume of RBC was associated with mortality for VV (SWD = -0.72, 95% CI: -1.18, -0.26, p = 0.002; I2 = 0.0%, Pheterogeneity = 0.642) and venoarterial (SWD = -0.95, 95% CI: -1.32, -0.57, p < .001) ECMO, but not when reported together (p = .067). The sensitivity analysis suggested the robustness of the results. CONCLUSION: When considering the total and daily volumes of RBC transfusion during ECMO, the patients who survived received smaller total and daily volumes of RBC transfusion. This meta-analysis suggests that RBC transfusion might be associated with a higher risk of mortality during ECMO.
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Internet of Things (IoT) applications have been increasingly developed. Authenticated key agreement (AKA) plays an essential role in secure communication in IoT applications. Without the PKI certificate and high time-complexity bilinear pairing operations, identity-based AKA (ID-AKA) protocols without pairings are more suitable for protecting the keys in IoT applications. In recent years, many pairing-free ID-AKA protocols have been proposed. Moreover, these protocols have some security flaws or relatively extensive computation and communication efficiency. Focusing on these problems, the security analyses of some recently proposed protocols have been provided first. We then proposed a family of eCK secure ID-AKA protocols without pairings to solve these security problems, which can be applied in IoT applications to guarantee communication security. Meanwhile, the security proofs of these proposed ID-AKA protocols are provided, which show they can hold provable eCK security. Some more efficient instantiations have been provided, which show the efficient performance of these proposed ID-AKA protocols. Moreover, comparisons with similar schemes have shown that these protocols have the least computation and communication efficiency at the same time.
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Zinc oxide nanorods were grown on an aluminum-doped zinc oxide seeds layer using the chemical bath deposition method. The effects of growth reaction time on the structural, optical, and photocatalytic properties of zinc oxide nanorods were investigated. It was clearly observed that the growth direction of zinc oxide nanorods were dependent on the crystallinity of the as-deposited aluminum-doped zinc oxide seed layer. The crystallinity of the obtained zinc oxide nanorods was improved with the increase in reaction times during the chemical bath deposition process. The mechanism of zinc oxide nanorod growth revealed that the growth rate of nanorods was influenced by the reaction times. With increasing reaction times, there were much more formed zinc oxide crystalline stacked growth along the c-axis orientation resulting in an increase in the length of nanorods. The longest nanorods and the high crystallinity were obtained from the zinc oxide nanorods grown within 5 h. The optical transmittance of all zinc oxide nanorods was greater than 70% in the visible region. Zinc oxide nanorods grown for 5 h showed the highest degradation efficiency of methyl red under ultraviolet light and had a high first-order degradation rate of 0.0051 min-1. The photocatalytic mechanism was revealed as well.
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Nanotubos , Óxido de Zinco , Óxido de Zinco/química , Tempo de Reação , Alumínio , Nanotubos/química , Raios UltravioletaRESUMO
In response to myocardial infarction (MI), quiescent cardiac fibroblasts differentiate into myofibroblasts mediating tissue repair. One of the most widely accepted markers of myofibroblast differentiation is the expression of Acta2 which encodes smooth muscle alpha-actin (SMαA) that is assembled into stress fibers. However, the requirement of Acta2/SMαA in the myofibroblast differentiation of cardiac fibroblasts and its role in post-MI cardiac repair remained unknown. To answer these questions, we generated a tamoxifen-inducible cardiac fibroblast-specific Acta2 knockout mouse line. Surprisingly, mice that lacked Acta2 in cardiac fibroblasts had a normal post-MI survival rate. Moreover, Acta2 deletion did not affect the function or histology of infarcted hearts. No difference was detected in the proliferation, migration, or contractility between WT and Acta2-null cardiac myofibroblasts. Acta2-null cardiac myofibroblasts had a normal total filamentous actin level and total actin level. Acta2 deletion caused a significant compensatory increase in the transcription level of non-Acta2 actin isoforms, especially Actg2 and Acta1. Moreover, in myofibroblasts, the transcription levels of cytoplasmic actin isoforms were significantly higher than those of muscle actin isoforms. In addition, we found that myocardin-related transcription factor-A is critical for myofibroblast differentiation but is not required for the compensatory effects of non-Acta2 isoforms. In conclusion, the Acta2 deletion does not prevent the myofibroblast differentiation of cardiac fibroblasts or affect the post-MI cardiac repair, and the increased expression and stress fiber formation of non-SMαA actin isoforms and the functional redundancy between actin isoforms are able to compensate for the loss of Acta2 in cardiac myofibroblasts.
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Actinas , Infarto do Miocárdio , Miofibroblastos , Actinas/genética , Actinas/metabolismo , Animais , Diferenciação Celular/genética , Fibroblastos/metabolismo , Camundongos , Infarto do Miocárdio/metabolismo , Miofibroblastos/metabolismo , Tamoxifeno/farmacologiaRESUMO
BACKGROUND: GenoLab M is a recently developed next-generation sequencing (NGS) platform from GeneMind Biosciences. To establish the performance of GenoLab M, we present the first report to benchmark and compare the WGS and WES sequencing data of the GenoLab M sequencer to NovaSeq 6000 and NextSeq 550 platform in various types of analysis. For WGS, thirty-fold sequencing from Illumina NovaSeq platform and processed by GATK pipeline is currently considered as the golden standard. Thus this dataset is generated as a benchmark reference in this study. RESULTS: GenoLab M showed an average of 94.62% of Q20 percentage for base quality, while the NovaSeq was slightly higher at 96.97%. However, GenoLab M outperformed NovaSeq or NextSeq at a duplication rate, suggesting more usable data after deduplication. For WGS short variant calling, GenoLab M showed significant accuracy improvement over the same depth dataset from NovaSeq, and reached similar accuracy to NovaSeq 33X dataset with 22x depth. For 100X WES, the F-score and Precision in GenoLab M were higher than NovaSeq or NextSeq, especially for InDel calling. CONCLUSIONS: GenoLab M is a promising NGS platform for high-performance WGS and WES applications. For WGS, 22X depth in the GenoLab M sequencing platform offers a cost-effective alternative to the current mainstream 33X depth on Illumina.