Real-time quantification of microRNAs by stem–loop RT–PCR

A novel microRNA (miRNA) quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis. Stem–loop RT primers are better than conventional ones in terms of RT efficiency and specificity. TaqMan miRNA assays are specific for mature miRNAs and discriminate among related miRNAs that differ by as little as one nucleotide. Furthermore, they are not affected by genomic DNA contamination. Precise quantification is achieved routinely with as little as 25 pg of total RNA for most miRNAs. In fact, the high sensitivity, specificity and precision of this method allows for direct analysis of a single cell without nucleic acid purification. Like standard TaqMan gene expression assays, TaqMan miRNA assays exhibit a dynamic range of seven orders of magnitude. Quantification of five miRNAs in seven mouse tissues showed variation from less than 10 to more than 30 000 copies per cell. This method enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases. Stem–loop RT–PCR can be used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs). Furthermore, the concept of stem–loop RT primer design could be applied in small RNA cloning and multiplex assays for better specificity and efficiency

RNA recognition by human TLR8 can lead to autoimmune inflammation.

Studies on the role of the RNA receptor TLR8 in inflammation have been limited by its different function in human versus rodents. We have generated multiple lines of transgenic mice expressing different levels of human TLR8. The high copy number chimeras were unable to pass germline; developed severe inflammation targeting the pancreas, salivary glands, and joints; and the severity of the specific phenotypes closely correlated with the huTLR8 expression levels. Mice with relatively low expression levels survived and bred successfully but had increased susceptibility to collagen-induced arthritis, and the levels of huTLR8 correlated with proinflammatory cytokines in the joints of the animals. At the cellular level, huTLR8 signaling exerted a DC-intrinsic effect leading to up-regulation of co-stimulatory molecules and subsequent T cell activation. A pathogenic role for TLR8 in human diseases was suggested by its increased expression in patients with systemic arthritis and the correlation of TLR8 expression with the elevation of IL-1β levels and disease status. We found that the consequence of self-recognition via TLR8 results in a constellation of diseases, strikingly distinct from those related to TLR7 signaling, and points to specific inflammatory diseases that may benefit from inhibition of TLR8 in humans

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

LiF as an Artificial SEI Layer to Enhance the High-Temperature Cycle Performance of Li4Ti5O12

Li4Ti6O12 (LTO) is a promising anode Material for electric vehicles (EVs) and electrochemical energy storage applications because of its safety, good rate capability, and long cycle life. At elevated temperature, such as 60 degrees C, it always shows poor cycle performance because of the instability between the electrode material and electrolyte, which may also lead to a serious gassing issue. In this article, a facile hydrothermal method is adopted to coat the LTO powder with a thin LiF layer, in which the LiF acts as an artificial, solid electrolyte interface (SEI) layer to prevent the direct contact of LTO and electrolyte, thus improving the high-temperature cycle performance. Electrochemical tests prove that the LiF coating layer has no influence on the kinetics at ambient temperature and greatly enhances the high-temperature cycle stability, and the LTO@LiF composite material keeps 87% of its initial discharge capacity in 300 1C cycles at 60 degrees C. Moreover, the LiF coating layer exhibits a special self-driven reforming process during the initial cycles, which makes it uniform and more effective at enhancing the stability between electrode/electrolyte interfaces.</p

Acute salinity and temperature challenges during early development of zebrafish: Differential gene expression of PTHs, PTHrPs and their receptors

Parathyroid hormone (pth), parathyroid hormone-related peptide (pthrp) and their receptors are involved in the regulation of calcium homeostasis in all vertebrates. To further understand the role of these genes in teleosts during development, we investigated the expression pattern of pth1, pth2, pthrp1, pthrp2, and their receptors pth1r, pth2r, pth3r, and their response to acute salinity and temperature challenge during early development of zebrafish, Danio rerio. The results revealed that pth1, pth2, pthrp1, pthrp2, pth1r, pth2r, pth3r were differentially expressed during early development, and pth1, pthrp1, pth1r, pth2r mRNA were detected from 0 hpf. pth2 and pth3r mRNA were detected after fertilization. Exposure of zebrafish embryos and larvae to acute osmotic (30) stress for 15 min failed to modify the expression levels of pthrp2 mRNA from levels in control fish. However, salinity challenge significantly (P < 0.01) modified pth1, pth2, pth3r at 3 dpf, pthrp1 (P < 0.01) and pth1r (P < 0.05) were both significantly modified at 5 dpf, and pth2r was significantly (P < 0.01) modified at 4 dpf and 5 dpf. Exposure of embryos and larvae to a cold (18 °C) stress generally up-regulated mRNA levels of pth1, pth2, pthrp1, pthrp2 and pth3r from 2 dpf to 5 dpf, while a hot (38 °C) stress generally down-regulated mRNA levels of these genes. After acute temperature challenge, expression levels of pth receptors were greatly influenced except pth2r. The results indicate that the contribution of pth1, pth2, pthrp1, pthrp2, pth1r, pth2r and pth3r genes to the stress response in zebrafish may be stressor-specific during early development. Overall, the results from this study provide a basis for further research into the developmental and stressor-specific role of pth1, pth2, pthrp1, pthrp2, pth1r, pth2r and pth3r in zebrafish

Teaching Machines to Extract Main Content for Machine Reading Comprehension

Machine reading comprehension, whose goal is to find answers from the candidate passages for a given question, has attracted a lot of research efforts in recent years. One of the key challenge in machine reading comprehension is how to identify the main content from a large, redundant, and overlapping set of candidate sentences. In this paper we propose to tackle the challenge with Markov Decision Process in which the main content identification is formalized as sequential decision making and each action corresponds to selecting a sentence. Policy gradient is used to learn the model parameters. Experimental results based on MSMARCO showed that the proposed model, called MC-MDP, can select high quality main contents and significantly improved the performances of answer span prediction

Fast and Sensitive Determination of Cadmium and Selenium in Rice by Direct Sampling Electrothermal Vaporization Inductively Coupled Plasma Mass Spectrometry

In this work, a direct solid sampling device based on modified graphite furnace electrothermal vaporization (GF-ETV) with inductively coupled plasma mass spectrometry (ICP-MS) was established for the simultaneous detection of trace selenium and cadmium in rice samples. A bypass gas was first designed in GF-ETV to improve the device&rsquo;s analytical sensitivity and precision. The ashing and vaporization conditions, the flow rates of the Ar carrier and the bypass gases of ICP-MS were all investigated. Under the optimized conditions, the limits of detection (LODs) for Se and Cd were 0.5 &mu;g kg&minus;1 and 0.16 &mu;g kg&minus;1, respectively; the relative standard deviations (RSDs) of repeated measurements were within 8% (n = 6). The recoveries of Cd and Se in rice samples were in the range of 89&ndash;112% compared with the microwave digestion ICP-MS method, indicating good accuracy and precision for the simultaneous detection of Se and Cd in rice matrix. The whole analysis time is &lt;3 min without the sample digestion process, fulfilling the fast detection of Se and Cd in rice samples to protect food safety

LiF as an Artificial SEI Layer to Enhance the High-Temperature Cycle Performance of Li₄Ti₅O₁₂

Li₄Ti₅O₁₂ (LTO) is a promising anode material for electric vehicles (EVs) and electrochemical energy storage applications because of its safety, good rate capability, and long cycle life. At elevated temperature, such as 60 °C, it always shows poor cycle performance because of the instability between the electrode material and electrolyte, which may also lead to a serious gassing issue. In this article, a facile hydrothermal method is adopted to coat the LTO powder with a thin LiF layer, in which the LiF acts as an artificial solid electrolyte interface (SEI) layer to prevent the direct contact of LTO and electrolyte, thus improving the high-temperature cycle performance. Electrochemical tests prove that the LiF coating layer has no influence on the kinetics at ambient temperature and greatly enhances the high-temperature cycle stability, and the LTO@LiF composite material keeps 87% of its initial discharge capacity in 300 1C cycles at 60 °C. Moreover, the LiF coating layer exhibits a special self-driven reforming process during the initial cycles, which makes it uniform and more effective at enhancing the stability between electrode/electrolyte interfaces

Fear Conditioning Downregulates Rac1 Activity in the Basolateral Amygdala Astrocytes to Facilitate the Formation of Fear Memory

Astrocytes are well known to scale synaptic structural and functional plasticity, while the role in learning and memory, such as conditioned fear memory, is poorly elucidated. Here, using pharmacological approach, we find that fluorocitrate (FC) significantly inhibits the acquisition of fear memory, suggesting that astrocyte activity is required for fear memory formation. We further demonstrate that fear conditioning downregulates astrocytic Rac1 activity in basolateral amygdala (BLA) in mice and promotes astrocyte structural plasticity. Ablation of astrocytic Rac1 in BLA promotes fear memory acquisition, while overexpression or constitutive activation of astrocytic Rac1 attenuates fear memory acquisition. Furthermore, temporal activation of Rac1 by photoactivatable Rac1 (Rac1-PA) induces structural alterations in astrocytes and in vivo activation of Rac1 in BLA astrocytes during fear conditioning attenuates the formation of fear memory. Taken together, our study demonstrates that fear conditioning-induced suppression of BLA astrocytic Rac1 activity, associated with astrocyte structural plasticity, is required for the formation of conditioned fear memory