Identification and Characterization of microRNAs from Peanut (Arachis hypogaea L.) by High-Throughput Sequencing

BACKGROUND: MicroRNAs (miRNAs) are noncoding RNAs of approximately 21 nt that regulate gene expression in plants post-transcriptionally by endonucleolytic cleavage or translational inhibition. miRNAs play essential roles in numerous developmental and physiological processes and many of them are conserved across species. Extensive studies of miRNAs have been done in a few model plants; however, less is known about the diversity of these regulatory RNAs in peanut (Arachis hypogaea L.), one of the most important oilseed crops cultivated worldwide. RESULTS: A library of small RNA from peanut was constructed for deep sequencing. In addition to 126 known miRNAs from 33 families, 25 novel peanut miRNAs were identified. The miRNA* sequences of four novel miRNAs were discovered, providing additional evidence for the existence of miRNAs. Twenty of the novel miRNAs were considered to be species-specific because no homolog has been found for other plant species. qRT-PCR was used to analyze the expression of seven miRNAs in different tissues and in seed at different developmental stages and some showed tissue- and/or growth stage-specific expression. Furthermore, potential targets of these putative miRNAs were predicted on the basis of the sequence homology search. CONCLUSIONS: We have identified large numbers of miRNAs and their related target genes through deep sequencing of a small RNA library. This study of the identification and characterization of miRNAs in peanut can initiate further study on peanut miRNA regulation mechanisms, and help toward a greater understanding of the important roles of miRNAs in peanut

Cross-Ocean Distribution of Rhodobacterales Bacteria as Primary Surface Colonizers in Temperate Coastal Marine Waters▿ †

Bacterial surface colonization is a universal adaptation strategy in aquatic environments. However, neither the identities of early colonizers nor the temporal changes in surface assemblages are well understood. To determine the identities of the most common bacterial primary colonizers and to assess the succession process, if any, of the bacterial assemblages during early stages of surface colonization in coastal water of the West Pacific Ocean, nonnutritive inert materials (glass, Plexiglas, and polyvinyl chloride) were employed as test surfaces and incubated in seawater off the Qingdao coast in the spring of 2005 for 24 and 72 h. Phylogenetic analysis of the 16S rRNA gene sequences amplified from the recovered surface-colonizing microbiota indicated that diverse bacteria colonized the submerged surfaces. Multivariate statistical cluster analyses indicated that the succession of early surface-colonizing bacterial assemblages followed sequential steps on all types of test surfaces. The Rhodobacterales, especially the marine Roseobacter clade members, formed the most common and dominant primary surface-colonizing bacterial group. Our current data, along with previous studies of the Atlantic coast, indicate that the Rhodobacterales bacteria are the dominant and ubiquitous primary surface colonizers in temperate coastal waters of the world and that microbial surface colonization follows a succession sequence. A conceptual model is proposed based on these findings, which may have important implications for understanding the structure, dynamics, and function of marine biofilms and for developing strategies to harness or control surface-associated microbial communities

Should they wait? Two children under 3 years old infected by HCV 1b successfully treated by ledipasvir/sofosbuvir: A report of two cases

Although direct-acting antivirals (DAAs) have notably increased the sustained virological response (SVR) rates in hepatitis C virus (HCV)-infected adolescent patients, the efficacy and safety for young children under 3 years old remain unclear. Currently, no guidelines recommend DAA therapy for this situation worldwide. Furthermore, the China National Medical Products Administration has not approved any DAA for treating children below 12 years old. Here, we described the characteristics of two children approximately 2 years old, who were infected by HCV genotype 1b and had significant clinical symptoms. Both received 12 weeks of ledipasvir/sofosbuvir (Case 1: 45.00 mg/200 mg per day, weight 17 kg; Case 2: 33.75 mg/150 mg per day, weight 12 kg). They achieved SVR at 12 weeks after treatment completion without obvious treatment-related adverse effects. Therefore, the safety and benefits of ledipasvir/sofosbuvir treatment in children under 3 years old seem to be confirmed. Our findings require further evaluation

Noninvasive Models to Assess Liver Inflammation and Fibrosis in Chronic HBV Infected Patients with Normal or Mildly Elevated Alanine Transaminase Levels: Which One Is Most Suitable?

The prevalence of substantial inflammation or fibrosis in treatment-naïve patients with chronic hepatitis B (CHB) and normal alanine transaminase (ALT) levels is high. A retrospective analysis was conducted on 559 consecutive patients with hepatitis B virus infection, who underwent liver biopsy, to investigate the value of noninvasive models based on routine serum markers for evaluating liver histology in CHB patients with normal or mildly elevated ALT levels and to provide treatment guidance. After comparing 55 models, we identified the top three models that exhibited excellent performance. The APGA model, based on the area under the receiver operating characteristic curve (AUROC), demonstrated a superior ability to evaluate significant (AUROC = 0.750) and advanced fibrosis (AUROC = 0.832) and demonstrated a good performance in assessing liver inflammation (AUROCs = 0.779 and 0.874 for stages G ≥ 2 and G ≥ 3, respectively). APGA also exhibited significant correlations with liver inflammation and fibrosis stage (correlation coefficients, 0.452 and 0.405, respectively (p < 0.001)). When the patients were stratified into groups based on HBeAg status and ALT level, APGA consistently outperformed the other 54 models. The other top two models, GAPI and XIE, also outperformed models based on other chronic hepatitis diseases. APGA may be the most suitable option for detecting liver fibrosis and inflammation in Chinese patients with CHB

An ultrasonic nanobubble-mediated PNP/fludarabine suicide gene system: A new approach for the treatment of hepatocellular carcinoma

<div><p>Objective</p><p>The purpose of this study is to generate an ultrasonic nanobubble (NB)-mediated purine nucleoside phosphorylase (PNP)/fludarabine suicide gene system for the treatment of human hepatocellular carcinoma (HCC).</p><p>Methods</p><p>NBs were prepared from a mixture the phospholipids 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphate (DPPA), perfluoropropane gas and other materials using the high shear dispersion method. NBs treated with ultrasound irradiation functioned as a gene-transfer system, and a self-constructed suicide gene expression plasmid, pcDNA3.1(+)/PNP, treated with fludarabine functioned as a therapeutic gene. This system was used to determine the cytotoxic effects of PNP/fludarabine on HepG2 cells and SMMC7721 cells.</p><p>Results</p><p><b>1.</b> NBs with a small diameter (208–416 nm) and at a high concentration and fine homogeneity were prepared under the optimal method. <b>2.</b> The pcDNA3.1(+)/PNP plasmid was efficiently transfected into HCC cells using ultrasonic NBs. <b>3.</b> At 0.75μg/ml fludarabine, PNP/fludarabine showed marked cytotoxic effects toward HepG2 and SMMC7721 cells. PNP/fludarabine achieved the same effect against both SMMC7721 and HepG2 cells but at a lower concentration of fludarabine for the latter. <b>4.</b> Bystander effects: a 10–20% decrease in the cell survival rate was observed when only 5–10% of transfected cells were PNP positive.</p><p>Conclusions</p><p>NBs constitute a non-toxic, stable and effective gene-delivery platform. The PNP/fludarabine suicide gene system inhibited the growth of HCC cells, induced HCC cell apoptosis, and caused a notable bystander effect at a low fludarabine concentration. This study establishes an important new method for miniaturizing microbubbles and improving a new NB-mediated approach for gene therapy of HCC.</p></div

MTT bioassay detection of the cellular growth inhibition effect of the PNP/fludarabine suicide gene system.

<p>MTT bioassay detection of the cellular growth inhibition effect of the PNP/fludarabine suicide gene system.</p

The “bystander” effect of the PNP/fludarabine suicide gene system.

<p>The cell survival rate decreased with the increasing ratio of transfected/non-transfected cells (<i>P</i><0.05).</p

Electrical tuning of radiative cooling at ambient conditions

Passive radiative cooling forms a sustainable means for cooling of objects through thermal radiation. Along with progress on static cooling systems, there is an emerging need for dynamic control to enable thermoregulation. Here, we demonstrate temperature regu-lation of devices at ambient pressure and temperature by electri-cally tuning their radiative cooling power. Our concept exploits the possibility to electrochemically tune the thermal emissivity and thereby cooling power of a conducting polymer, which enabled reversible control of device temperatures of around 0.25 degrees C at ambient conditions in a sky simulator. Besides tuneable radiative cooling by exposure to the sky, the concept could also contribute to reduced needs for indoor climate control by enabling dynamic control of thermal energy flows between indoor objects, such as be-tween people and walls.Funding Agencies|Knut and Alice Wallenberg Foundation; Linkoping University; Wallenberg Wood Science Center; Swedish government; Strategic Research Area in Materials Science on Functional Materials at Linkoping University (faculty grant SFO-Mat-LiU) [2009 00971]; Swedish Armed Forces Research; Swedish Research Council [2020-00287]; Wallenberg Academy [2019.0163]</p

Differences in GFP transfection efficiencies of NBs and liposomes (×200) and GFP transfection with or without ultrasound irradiation (×200).

<p>a. 8 μg of plasmid; b. 8 μg of plasmid and ultrasound irradiation; c. NBs and 8 μg of plasmid; d. NBs, 8 μg of plasmid and ultrasound irradiation; e. liposomes and 8 μg of plasmid; f. liposomes, 8 μg of plasmid and ultrasound irradiation; g. NBs and 4 μg of plasmid; h. liposomes and 4 μg of plasmid; i. FCM assessment of GFP expression in transfected cells with or without ultrasound irradiation; expression in group d is significantly higher than in other groups (*<i>P</i><0.05).</p