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  • Latest Articles

    Open AccessReview
    Controlling the Oxygen Electrocatalysis on Perovskite and Layered Oxide Thin Films for Solid Oxide Fuel Cell Cathodes
    Appl. Sci. 2019, 9(5), 1030; https://doi.org/10.3390/app9051030 (registering DOI) -
    Abstract
    Achieving the fast oxygen reduction reaction (ORR) kinetics at the cathode of solid oxide fuel cells (SOFCs) is indispensable to enhance the efficiency of SOFCs at intermediate temperatures. Mixed ionic and electronic conducting (MIEC) oxides such as ABO3 perovskites and Ruddlesden-Popper (RP) [...] Read more.
    Achieving the fast oxygen reduction reaction (ORR) kinetics at the cathode of solid oxide fuel cells (SOFCs) is indispensable to enhance the efficiency of SOFCs at intermediate temperatures. Mixed ionic and electronic conducting (MIEC) oxides such as ABO3 perovskites and Ruddlesden-Popper (RP) oxides (A2BO4) have been widely used as promising cathode materials owing to their attractive physicochemical properties. In particular, oxides in forms of thin films and heterostructures have enabled significant enhancement in the ORR activity. Therefore, we aim to give a comprehensive overview on the recent development of thin film cathodes of SOFCs. We discuss important advances in ABO3 and RP oxide thin film cathodes for SOFCs. Our attention is also paid to the influence of oxide heterostructure interfaces on the ORR activity of SOFC cathodes. Full article
    Open AccessArticle
    Flood Resilience of Critical Infrastructure: Approach and Method Applied to Fort Lauderdale, Florida
    Water 2019, 11(3), 517; https://doi.org/10.3390/w11030517 (registering DOI) -
    Abstract
    In order to increase the flood resilience of cities (i.e., the ability to cope with flood hazards), it is also crucial to make critical infrastructure functions resilient, since these are essential for urban society. Cities are complex systems with many actors of different [...] Read more.
    In order to increase the flood resilience of cities (i.e., the ability to cope with flood hazards), it is also crucial to make critical infrastructure functions resilient, since these are essential for urban society. Cities are complex systems with many actors of different disciplines and many interdependent critical infrastructure networks and functions. Common flood risk analysis techniques provide useful information but are not sufficient to obtain a complete overview of the effects of flooding and potential measures to increase flood resilience related to critical infrastructure networks. Therefore, a more comprehensive approach is needed which helps accessing knowledge of actors in a structured way. Fort Lauderdale, Florida, United States has suffered from flood impacts, especially from disruptions in critical infrastructure. This paper shows how shared insight among different sectors and stakeholders into critical infrastructure resilience and potential resilience-enhancing measures was obtained using input from these actors. It also provides a first quantitative indication of resilience, indicated by the potential disruption due to floods and the effect of measures on resilience. The paper contributes to the existing literature on resilience specifically by considering the duration of disruption, the inclusion of critical infrastructure disruption in flood impact analysis, and the step from resilience quantification to measures. Full article
    Open AccessReview
    Novel Therapeutics for Epstein–Barr Virus
    Molecules 2019, 24(5), 997; https://doi.org/10.3390/molecules24050997 (registering DOI) -
    Abstract
    Epstein–Barr virus (EBV) is a human γ-herpesvirus that infects up to 95% of the adult population. Primary EBV infection usually occurs during childhood and is generally asymptomatic, though the virus can cause infectious mononucleosis in 35–50% of the cases when infection occurs later [...] Read more.
    Epstein–Barr virus (EBV) is a human γ-herpesvirus that infects up to 95% of the adult population. Primary EBV infection usually occurs during childhood and is generally asymptomatic, though the virus can cause infectious mononucleosis in 35–50% of the cases when infection occurs later in life. EBV infects mainly B-cells and epithelial cells, establishing latency in resting memory B-cells and possibly also in epithelial cells. EBV is recognized as an oncogenic virus but in immunocompetent hosts, EBV reactivation is controlled by the immune response preventing transformation in vivo. Under immunosuppression, regardless of the cause, the immune system can lose control of EBV replication, which may result in the appearance of neoplasms. The primary malignancies related to EBV are B-cell lymphomas and nasopharyngeal carcinoma, which reflects the primary cell targets of viral infection in vivo. Although a number of antivirals were proven to inhibit EBV replication in vitro, they had limited success in the clinic and to date no antiviral drug has been approved for the treatment of EBV infections. We review here the antiviral drugs that have been evaluated in the clinic to treat EBV infections and discuss novel molecules with anti-EBV activity under investigation as well as new strategies to treat EBV-related diseases. Full article
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    <p>EBV life cycle, latency stages and derived lymphomas. The viral life cycle includes at least five different stages (virus entry, infection, proliferation, differentiation and persistence), and four of them are associated with EBV diseases. The virus is transmitted through the saliva and infects na?ve B-cells in the oropharyngeal mucosa. During primary infection, EBV-infected na?ve B-cells express the entire latency gene complex (10 proteins: EBV nuclear antigens (EBNAs), latent membrane protein (LMPs)) as well as EBV-encoded small RNAs (EBERs) and microRNAs. This is called type III latency and this form of latency activates the resting B-cells and drives them to proliferation and transformation. However, these cells are highly immunogenic and are rapidly eliminated by EBV-specific T cells. The virus is able to survive in B-cells because it downregulates its immunogenic proteins. EBV mimics antigen driven B-cell responses and similar to antigen-stimulated blasts, the EBV-infected B-cells enter the follicles, expand, and form germinal centers where they express only three viral proteins (type II latency). Finally, they exit the lymph node expressing only a single viral protein (EBNA1, which ensures that the viral genome divides with the cellular genome) (type I latency). The entry of EBV-infected cells into the peripheral blood results in the shutdown of all viral genes encoding for proteins; this is called latency 0 or latency program where no viral proteins are expressed. Resting memory cells, in which the virus is quiescent, are not attacked by the host immune system and are likely the sites of long-term persistence. Memory B cells occasionally divide to maintain stable number of cells and when a cell that is carrying the virus divides, the viral EBNA1 protein is expressed to allow the viral genome to replicate along with the cell. Memory B-cells may also undergo terminal differentiation into plasma cells and secrete antibodies. If such a cell contains the virus, the EBV lytic program is activated and the infectious virus released from the plasma cells can infect epithelial cells, where the virus can replicate and be shed at high amounts and then be transmitted to other hosts. With the exception of latency type 0, each latency state is found in specific types of EBV-associated malignancies.</p>
    Full article ">Figure 2
    <p>(<bold>a</bold>) Patterns of gene expression during EBV latency. The majority of the endemic BL presents a latency I type and carry a wild-type transformation-competent EBV genome and express only the Epstein–Barr nuclear antigen 1 (EBNA1) from the <italic>EBNA1</italic>-specific latent promoter Qp, non-coding EBERs (Epstein–Barr virus-encoded small RNAs) and several microRNAs (miRNAs). Around 15% of BL endemic tumors, the so called Wp-restricted BLs, carry an <italic>EBNA2</italic> gene-deleted genome and express EBNA1, -3A, -3B, and-3C and the viral Bcl2 homologue BHRF1 from the Wp latent promoter [<xref ref-type="bibr" rid="B2-molecules-24-00997">2</xref>,<xref ref-type="bibr" rid="B6-molecules-24-00997">6</xref>]. * The EBNA-LP gene is partially deleted in the Wp-restricted latency. A major type of latency in EBV-associated malignancies is latency II, in which the latent membrane proteins LMP1, LMP2A, and LMP2B are expressed in addition to the Latency I genes. The entire EBV latency gene complex, which consists of several EBNA proteins, LMP1, LMP2A, LMP2B, EBERs, and miRNAs are expressed in the type III latency. (<bold>b</bold>) The cellular genetic alterations and/or co-infections are known to occur in the different types of EBV-associated malignancies. PEL: primary effusion lymphoma; HL: Hodgkin lymphoma; BL: Burkitt lymphoma; NHL: non-Hodgkin lymphoma; PTLD: post-transplant lymphoproliferative disorder; NPC: nasopharyngeal carcinoma; GC: gastric carcinoma.</p>
    Full article ">
    Open AccessArticle
    Thoracic Paravertebral Block with Adjuvant Dexmedetomidine in Video-Assisted Thoracoscopic Surgery: A Randomized, Double-Blind Study
    J. Clin. Med. 2019, 8(3), 352; https://doi.org/10.3390/jcm8030352 (registering DOI) -
    Abstract
    Background: The addition of the adjuvant dexmedetomidine to a nerve block improves the quality of the block and reduces perioperative opioid consumption. The aim of this study was to assess the effect of dexmedetomidine as an adjuvant for the thoracic paravertebral block (TPVB) [...] Read more.
    Background: The addition of the adjuvant dexmedetomidine to a nerve block improves the quality of the block and reduces perioperative opioid consumption. The aim of this study was to assess the effect of dexmedetomidine as an adjuvant for the thoracic paravertebral block (TPVB) in postoperative pain control after video-assisted thoracoscopic surgery (VATS). Methods: Sixty-six males, aged 15–40 years, with spontaneous pneumothorax scheduled for VATS wedge resection were enrolled. Following surgery, ultrasound-guided TPVB was performed on the T3 and T5 levels with 30 mL of 0.5% ropivacaine, plus adjuvant dexmedetomidine 50 μg or normal saline. The primary outcome was cumulative fentanyl consumption at 24 h. Pain severity, the requirement for additional rescue analgesics, hemodynamic variations, and side effects were also evaluated. Results: Median postoperative cumulative fentanyl consumption at 24 h was significantly lower in the dexmedetomidine group (122.6 (interquartile range (IQR) 94.5–268.0) μg vs. 348.1 (IQR, 192.8–459.2) μg, p-value = 0.001) with a Hodges–Lehman median difference between groups of 86.2 (95% confidence interval (CI), 4.2–156.4) mg. Coughing numeric rating scale (NRS) was lower in the dexmedetomidine group at postoperative 2, 4, 8, and 24 h. However, resting NRS differed significantly only after 4 h postoperative. Conclusions: Dexmedetomidine as an adjunct in TPVB provided effective pain relief and significantly reduced opioid requirement in VATS. Full article
    Figures

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    Figure 1
    <p>The Consolidated Standards of Reporting Trials (CONSORT) flow diagram of study participants. TPVB: thoracic paravertebral block.</p>
    Full article ">Figure 2
    <p>Cumulative fentanyl consumption over time in both groups. Data are expressed as median (interquartile range). * <italic>p</italic> &lt; 0.05. ● outlier (any data point more than 1.5 interquartile ranges below the first quartile or above the third quartile).</p>
    Full article ">Figure 3
    <p>Pain score over time while (<bold>a</bold>) resting and (<bold>b</bold>) coughing. Data are expressed as median (interquartile range). * <italic>p</italic> &lt; 0.05. ● outlier (any data point more than 1.5 interquartile ranges below the first quartile or above the third quartile).</p>
    Full article ">Figure 4
    <p>Changes in heart rate over time in the post-anesthesia care unit. The changes over time differed significantly in the two groups (<italic>p</italic> = 0.002). * <italic>p</italic> &lt; 0.013 using Bonferroni’s correction for multiple comparisons. PACU: post-anesthesia care unit; TPVB: thoracic paravertebral block.</p>
    Full article ">
    Open AccessArticle
    Differential Proteomics Reveals miR-155 as a Novel Indicator of Liver and Spleen Pathology in the Symptomatic Niemann-Pick Disease, Type C1 Mouse Model
    Molecules 2019, 24(5), 994; https://doi.org/10.3390/molecules24050994 (registering DOI) -
    Abstract
    Niemann-Pick disease, type C1 (NPC1) is a rare, autosomal recessive, lipid storage disorder caused by mutations in NPC1. As a result, there is accumulation of unesterified cholesterol and sphingolipids in the late endosomal/lysosomal system. Clinically, patients can present with splenomegaly and hepatomegaly. 秒速赛车是哪里的开奖: [...] Read more.
    Niemann-Pick disease, type C1 (NPC1) is a rare, autosomal recessive, lipid storage disorder caused by mutations in NPC1. As a result, there is accumulation of unesterified cholesterol and sphingolipids in the late endosomal/lysosomal system. Clinically, patients can present with splenomegaly and hepatomegaly. In the current study, we analyzed the differential proteome of the spleen in symptomatic Npc1−/− mice to complement previous studies focused on the differential proteome of the liver, and then evaluated biomolecules that may serve as tissue biomarkers. The proteomic analysis revealed altered pathways in NPC1 representing different functional categories including heme synthesis, cellular regulation and phosphoinositide metabolism in both tissues. Differential proteins included several activators of the ubiquitous and critical protein, Akt, a major kinase involved in multiple cellular processes. Evaluation of Akt revealed decreased expression in both the liver and spleen tissues of symptomatic Npc1−/− mice. Upstream regulation analysis also suggested that miR-155 may modulate the differences of known downstream protein targets observed in our dataset. Upon evaluation of miR-155, we observed an increased expression in the liver and decreased expression in the spleen of symptomatic Npc1−/− mice. Here, we propose that miR-155 may be a novel indicator of spleen and liver pathology in NPC1. Full article
    Figures

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    Figure 1
    <p>Comparative analysis of the differential proteome of the spleen and liver of 11-week <italic>Npc1</italic><sup>?/?</sup> mice. Differential proteins from the mass spectrometry analysis of the spleen (<bold>A</bold>) and liver (<bold>B</bold>) of <italic>Npc1</italic><sup>?/?</sup> mice where analyzed for downregulated and upregulated proteins. Evaluation of the proteome of the spleen revealed that 18.6% of the measured proteome was found to be downregulated while 2.6% was upregulated. The differential proteome of the liver was evaluated and revealed that 15.8% of the measured proteome was found to be downregulated while 7.9% was upregulated. Venn diagram analysis of the downregulated (<bold>C</bold>) and upregulated (<bold>D</bold>) proteins was performed to determine common altered proteins.</p>
    Full article ">Figure 2
    <p>Pathway analysis of the differential proteome of the spleen and liver of 11-week <italic>Npc1</italic><sup>?/?</sup> mice. Differential proteins from the mass spectrometry analysis of the spleen (<bold>A</bold>) and liver (<bold>B</bold>) of <italic>Npc1</italic><sup>?/?</sup> mice were enriched for the top 15 pathways. Significance was determined using a Right-tailed Fisher’s exact test to determine probability of pathways from the Ingenuity Pathway Analysis Knowledge Base Library to those most significantly enriched.</p>
    Full article ">Figure 3
    <p>Functional analysis of the differential proteome of the spleen and liver of 11-week <italic>Npc1</italic><sup>?/?</sup> mice. Differential proteins from the mass spectrometry analysis of the spleen (<bold>A</bold>) and liver (<bold>B</bold>) of <italic>Npc1</italic><sup>?/?</sup> mice where enriched for top biological functions. Significance was determined using a Right-tailed Fisher’s exact test to determine probability of pathways from the Ingenuity Pathway Analysis Knowledge Base Library to those most significantly enriched.</p>
    Full article ">Figure 4
    <p>Upstream regulator analysis of the differential proteome of the spleen from 11-week <italic>Npc1</italic><sup>?/?</sup> mice. Predicted upstream regulator analysis from 11-week <italic>Npc1</italic><sup>?/?</sup> mice suggest that Tcl1a is a common regulator of various deferential proteins of the spleen. Assignment was based on the Ingenuity Pathway Analysis Knowledge Base Library. GPNMB = Transmembrane glycoprotein NMB, GUSB = Beta-glucuronidase, ITGAM = Integrin alpha-M, MMP9 = Matrix metalloproteinase-9, MPEG1 = Macrophage-expressed gene 1 protein, Ngp = Nucleolar GTP-binding protein 1, SNX6 = Sorting nexin = 6, C1QC = Complement C1q subcomponent subunit C, CELA2A = Chymotrypsin-like elastase family member 2A, CTSD = Cathepsin D and CTSE = Cathepsin E.</p>
    Full article ">Figure 5
    <p>Western blot analysis of Akt in spleen and liver tissue of 11-week <italic>Npc1<sup>+/+</sup></italic> and <italic>Npc1</italic><sup>?/?</sup> mice. Protein lysates from the spleen and liver of 11-week <italic>Npc1<sup>+/+</sup></italic> and <italic>Npc1</italic><sup>?/?</sup> mice (N = 3 each genotype) were subject to electrophoresis and Western blotting. Data analysis revealed decreased expression of Akt in both the (<bold>A</bold>) spleen and (<bold>B</bold>) liver of <italic>Npc1</italic><sup>?/?</sup> mice. Data is reported as relative to Gapdh.</p>
    Full article ">Figure 6
    <p>miR-155 analysis of spleen and liver tissue from 9-week <italic>Npc1<sup>+/+</sup></italic> and <italic>Npc1</italic><sup>?/?</sup> mice. miR-155 was selected for evaluation by quantitative real-time PCR in spleen and liver tissue from <italic>Npc1<sup>+/+</sup></italic> and <italic>Npc1</italic><sup>?/?</sup> mice (N = 5 for each genotype). The levels of miR-155 were significantly decreased in the (<bold>A</bold>) spleen and increased in the (<bold>B</bold>) liver of <italic>Npc1</italic><sup>?/?</sup> mice. The expression level of the miR-155 in each tissue was normalized to endogenous U6 snRNA.</p>
    Full article ">
    Open AccessArticle
    Toward Content-Based Hyperspectral Remote Sensing Image Retrieval (CB-HRSIR): A Preliminary Study Based on Spectral Sensitivity Functions
    Remote Sens. 2019, 11(5), 600; https://doi.org/10.3390/rs11050600 (registering DOI) -
    Abstract
    With the emergence of huge volumes of high-resolution Hyperspectral Images (HSI)
    produced by different types of imaging sensors, analyzing and retrieving these images require
    effective image description and quantification techniques. Compared to remote sensing RGB images,
    HSI data contain hundreds of spectral bands [...] Read more.
    With the emergence of huge volumes of high-resolution Hyperspectral Images (HSI)
    produced by different types of imaging sensors, analyzing and retrieving these images require
    effective image description and quantification techniques. Compared to remote sensing RGB images,
    HSI data contain hundreds of spectral bands (varying from the visible to the infrared ranges) allowing
    profile materials and organisms that only hyperspectral sensors can provide. In this article, we study
    the importance of spectral sensitivity functions in constructing discriminative representation of
    hyperspectral images. The main goal of such representation is to improve image content recognition
    by focusing the processing on only the most relevant spectral channels. The underlying hypothesis
    is that for a given category, the content of each image is better extracted through a specific set of
    spectral sensitivity functions. Those spectral sensitivity functions are evaluated in a Content-Based
    Image Retrieval (CBIR) framework. In this work, we propose a new HSI dataset for the remote
    sensing community, specifically designed for Hyperspectral remote sensing retrieval and classification.
    Exhaustive experiments have been conducted on this dataset and on a literature dataset. Obtained
    retrieval results prove that the physical measurements and optical properties of the scene contained
    in the HSI contribute in an accurate image content description than the information provided by the
    RGB image presentation. Full article
    Figures

    Graphical abstract

    Graphical abstract
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    Open AccessArticle
    Food Intake during School Lunch Is Better Explained by Objectively Measured Eating Behaviors than by Subjectively Rated Food Taste and Fullness: A Cross-Sectional Study
    Nutrients 2019, 11(3), 597; https://doi.org/10.3390/nu11030597 (registering DOI) -
    Abstract
    School lunches contribute significantly to students’ food intake (FI) and are important to their long-term health. Objective quantification of FI is needed in this context. The primary aim of this study was to investigate how much eating rate (g/min), number of food additions, [...] Read more.
    School lunches contribute significantly to students’ food intake (FI) and are important to their long-term health. Objective quantification of FI is needed in this context. The primary aim of this study was to investigate how much eating rate (g/min), number of food additions, number of spoonfuls, change in fullness, food taste, body mass index (BMI), and sex explain variations in school lunch FI. The secondary aim was to assess the reliability of repeated FI measures. One hundred and three (60 females) students (15–18 years old) were monitored while eating lunch in their normal school canteen environment, following their usual school schedules. A subgroup of students (n = 50) participated in a repeated lunch (~3 months later). Linear regression was used to explain variations in FI. The reliability of repeated FI measurements was assessed by change in mean, coefficient of variation (CV), and intraclass correlation (ICC). The regression model was significant and explained 76.6% of the variation in FI. Eating rate was the strongest explanatory variable, followed by spoonfuls, sex, food additions, food taste, BMI, and change in fullness. All explanatory variables were significant in the model except BMI and change in fullness. No systematic bias was observed in FI (−7.5 g (95% CI = −43.1–28 g)) while individual students changed their FI from −417 to +349 g in the repeated meal (CV 26.1% (95% CI = 21.4–33.5%), ICC 0.74 (95% CI = 0.58–0.84)). The results highlight the importance of objective eating behaviors for explaining FI in a school lunch setting. Furthermore, our methods show promise for large-scale quantification of objectively measured FI and eating behaviors in schools. Full article
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    <p>The lunchroom where the participating students ate their lunches.</p>
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    <p>Study diagram.</p>
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    <p>Placement of the cameras recording students during their lunches.</p>
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    <p>Food choices among a sample of the included students.</p>
    Full article ">Figure 5
    <p>Individual differences in recorded food intake (grams) between two lunches with identical food choices in a Swedish high school (<italic>n</italic> = 50). Black bars: female participants; Grey bars: male participants.</p>
    Full article ">
    Open AccessArticle
    Equations for Deep Water Counter Streaming Waves and New Integrals of Motion
    Fluids 2019, 4(1), 47; https://doi.org/10.3390/fluids4010047 (registering DOI) -
    Abstract
    The waves on a free surface of 2D deep water can be split into two groups: the waves moving to the right, and the waves moving to the left. A specific feature of the four-wave interactions of water waves allows to describe the [...] Read more.
    The waves on a free surface of 2D deep water can be split into two groups: the waves moving to the right, and the waves moving to the left. A specific feature of the four-wave interactions of water waves allows to describe the evolution of the two groups as a system of two equations. The fundamental consequence of this decomposition is the conservation of the “number of waves” in each particular group. The envelope approximation for the waves in each group of counter streaming waves is obtained. Full article
    Figures

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    <p>Two narrow bands spectra for two counter-streaming waves.</p>
    Full article ">
    Open AccessArticle
    Development of DNA Aptamers to Native EpCAM for Isolation of Lung Circulating Tumor Cells from Human Blood
    Cancers 2019, 11(3), 351; https://doi.org/10.3390/cancers11030351 (registering DOI) -
    Abstract
    We selected DNA aptamers to the epithelial cell adhesion molecule (EpCAM) expressed on primary lung cancer cells isolated from the tumors of patients with non-small cell lung cancer using competitive displacement of aptamers from EpCAM by a corresponding antibody. The resulting aptamers clones [...] Read more.
    We selected DNA aptamers to the epithelial cell adhesion molecule (EpCAM) expressed on primary lung cancer cells isolated from the tumors of patients with non-small cell lung cancer using competitive displacement of aptamers from EpCAM by a corresponding antibody. The resulting aptamers clones showed good nanomolar affinity to EpCAM-positive lung cancer cells. Confocal microscopy imaging and spectral profiling of lung cancer tissues confirmed the same protein target for the aptamers and anti-EpCAM antibodies. Furthermore, the resulted aptamers were successfully applied for isolation and detection of circulating tumor cells in clinical samples of peripheral blood of lung cancer patients. Full article
    Figures

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    Figure 1
    <p>The scheme of DNA aptamer selection using aptamer displacement via antibody. The first several rounds include only positive selection and start with the incubation of the ssDNA library or aptamer pools with receptor positive cells, followed by partitioning unbound DNA, and amplifying bound DNA with symmetric and asymmetric polymerase chain reaction (PCR). In the next rounds, positive rounds alternate with antibody displacement steps and include the incubation of aptamers with the receptor positive cells, washing, the displacement of the bound aptamers by antibodies (Ab), and the following amplification of free aptamers.</p>
    Full article ">Figure 2
    <p>Binding evaluation of aptamer pools. Flow cytometry of lung cancer (LC) cells incubated with pools of 6–9th rounds of aptamer selection against EpCAM at the first step and displaced by EpCAM antibodies at the second step in comparison with LC cells alone.</p>
    Full article ">Figure 3
    <p>Competitive displacement of aptamers with antibodies. (<bold>A</bold>) Flow cytometry of LC cells and LC cells preincubated with Cy-3 labeled anti-EpCAM or anti-α-Tubulin antibodies. (<bold>B</bold>) Flow cytometry of LC cells (red), LC cells preincubated with 6-carboxyfluorescein (FAM)-labeled EPCAM-APT-01, EPCAM-APT-02 or oligonucleotide (AG)<sub>40</sub> before (green) and after (blue) replacement by Cy-3 labeled anti-EpCAM or anti-α-Tubulin antibodies.</p>
    Full article ">Figure 4
    <p>Aptamer affinity curves. The percentage of bound LC cells measured by flow cytometry versus concentrations of EPCAM-APT-01 or EPCAM-APT-02.</p>
    Full article ">Figure 5
    <p>Co-staining aptamers and antibodies. Confocal microscopy of different regions of two squamous LC tissue sections stained with Alexa-Fluor 405-labeled anti-EpCAM antibodies and Cy-5-labeled aptamers EPCAM-APT-01 (<bold>A</bold>) and EPCAM-APT-02 (<bold>B</bold>). (<bold>A1</bold>,<bold>A5</bold>,<bold>B1</bold>,<bold>B5</bold>)—fluorescence of Cy-5-labeled aptamers, (<bold>A2</bold>,<bold>A6</bold>,<bold>B2</bold>,<bold>B6</bold>)—fluorescence of Alexa 405-labeled anti-EpCAM antibodies, (<bold>A3</bold>,<bold>A7</bold>,<bold>B3</bold>,<bold>B7</bold>)—overlays, (<bold>A4</bold>,<bold>A8</bold>,<bold>B4</bold>,<bold>B8</bold>)—overlaid fluorescence intensity spectra from the marked (<bold>A3</bold>,<bold>A7</bold>,<bold>B3</bold>,<bold>B7</bold>)—regions.</p>
    Full article ">Figure 5 Cont.
    <p>Co-staining aptamers and antibodies. Confocal microscopy of different regions of two squamous LC tissue sections stained with Alexa-Fluor 405-labeled anti-EpCAM antibodies and Cy-5-labeled aptamers EPCAM-APT-01 (<bold>A</bold>) and EPCAM-APT-02 (<bold>B</bold>). (<bold>A1</bold>,<bold>A5</bold>,<bold>B1</bold>,<bold>B5</bold>)—fluorescence of Cy-5-labeled aptamers, (<bold>A2</bold>,<bold>A6</bold>,<bold>B2</bold>,<bold>B6</bold>)—fluorescence of Alexa 405-labeled anti-EpCAM antibodies, (<bold>A3</bold>,<bold>A7</bold>,<bold>B3</bold>,<bold>B7</bold>)—overlays, (<bold>A4</bold>,<bold>A8</bold>,<bold>B4</bold>,<bold>B8</bold>)—overlaid fluorescence intensity spectra from the marked (<bold>A3</bold>,<bold>A7</bold>,<bold>B3</bold>,<bold>B7</bold>)—regions.</p>
    Full article ">Figure 6
    <p>Aptamer-facilitated isolation of circulating tumor cells (CTCs). CTCs were isolated from the blood of two LC patients: ID#101 (<bold>A1</bold>–<bold>A3</bold>) and ID#113 (<bold>B1</bold>–<bold>B3</bold>,<bold>C1</bold>–<bold>C3</bold>), using biotinylated aptamers EPCAM-APT-01 and EPCAM-APT-02 and then stained with the same fluorescent aptamers.</p>
    Full article ">
    Open AccessReview
    Autophagy and Noroviruses
    Viruses 2019, 11(3), 244; https://doi.org/10.3390/v11030244 (registering DOI) -
    Abstract
    Autophagy is an essential cellular process by which a cell degrades materials within its cytoplasm. Intracellular pathogens like viruses must deal with autophagy, either positively or negatively, for their own survival and replication. For some viruses, autophagy can even play proviral roles, helping [...] Read more.
    Autophagy is an essential cellular process by which a cell degrades materials within its cytoplasm. Intracellular pathogens like viruses must deal with autophagy, either positively or negatively, for their own survival and replication. For some viruses, autophagy can even play proviral roles, helping their replication or dissemination. For other viruses, including noroviruses, the exact role of autophagy is more complex. This short review seeks to summarize the known interactions between autophagy, autophagy proteins and norovirus, and to address remaining questions relevant to these interactions. Full article
    Open AccessTechnical Note
    Water Footprint and Water Pinch Analysis in Ethanol Industrial Production for Water Management
    Water 2019, 11(3), 518; https://doi.org/10.3390/w11030518 (registering DOI) -
    Abstract
    Fuel ethanol is considered to be a clean alternative fuel to meet increasing energy demands and mitigate environmental pollution. Faced with challenges in terms of energy security and environmental pollution, China is vigorously developing fuel ethanol. However, ethanol-manufacturing is a water-intensive industry; it [...] Read more.
    Fuel ethanol is considered to be a clean alternative fuel to meet increasing energy demands and mitigate environmental pollution. Faced with challenges in terms of energy security and environmental pollution, China is vigorously developing fuel ethanol. However, ethanol-manufacturing is a water-intensive industry; it consumes large volumes of fresh water and generates a corresponding amount of waste water. Expansion of this industry can reduce water quality and cause water stress. This study aims to combine the water footprint (WF) with a water pinch analysis technique to manage water consumption and sewage discharge systematically in an ethanol plant. A well-operated cassava ethanol plant in China was chosen as a case study. The WF of industrial ethanol production was evaluated. The total WF was 17.08 L/L ethanol, comprised of a 7.69 L blue water footprint (BWF), and a 9.39 L gray water footprint (GWF). The direct WF was 16.38 L/L ethanol, and the indirect WF was 0.70 L/L ethanol. Thereafter, a water pinch analysis was conducted, and the optimal direct water reuse scheme was studied. After the water network was optimized, the BWF was reduced by 0.98 L/L ethanol, while the GWF was reduced by 1.47 L/L ethanol. These results indicate that the combined use of WF and pinch analysis can provide the starch-based ethanol industry with an effective tool to improve its water management. Full article
    Open AccessCommentary
    Viroids as Companions of a Professional Career
    Viruses 2019, 11(3), 245; https://doi.org/10.3390/v11030245 (registering DOI) -
    Abstract
    Since the early 1970s when “virus-like” agents were considered as the cause of two diseases (potato spindle tuber and citrus exocortis), their study and further characterization have been linked to the development and use of molecular biology tools. Sucrose density gradient centrifugation and [...] Read more.
    Since the early 1970s when “virus-like” agents were considered as the cause of two diseases (potato spindle tuber and citrus exocortis), their study and further characterization have been linked to the development and use of molecular biology tools. Sucrose density gradient centrifugation and polyacrylamide gel electrophoresis (PAGE) played a critical role in the pioneering studies of PSTVd and citrus exocortis viroid (CEVd). This was later modified by using other PAGEs (sequential PAGE, return PAGE, two-dimensional PAGE), and/or different staining methods (ethidium bromide, silver nitrate, etc.). Since then, disease-causing agents suspected to be viroids were usually subjected to a number of tests to define their: (i) Molecular nature (RNA or DNA; single stranded or double stranded; circular or linear RNA); (ii) molecular weight; (iii) secondary and tertiary structure. Further biological assays are also essential to establish the relationship of a viroid with plant disease and to fulfill Koch’s postulates. Full article
    Figures

    Figure 1

    Figure 1
    <p>Analysis of different inoculated hosts by polyacrilamide gel electrophoresis (PAGE) and ethidium bromide staining. A characteristic band corresponding to a viroid RNA was stained in samples (1–3) from <italic>Gynura aurantiaca</italic> (<bold>A</bold>), tomato (<bold>B</bold>), and Etrog citron showing severe symptoms, whereas the same band was not observed in samples (4,5) from Etrog citron plants displaying mild and moderate symptoms (<bold>D</bold>–<bold>F</bold>).</p>
    Full article ">Figure 2
    <p>Analysis of inoculated Etrog citron plants by sPAGE and silver staining. Only plants showing severe symptoms (<bold>A</bold>) contained a band with the characteristic mobility of citrus exocortis viroid (CEVd) (1). Analysis of Etrog citron plants displaying mild and moderate symptoms (<bold>B</bold>–<bold>D</bold>) revealed bands with electrophoretic mobility different from CEVd (2–7). Virtually all plants contained a mixture of viroid-like RNAs that had to be further characterized.</p>
    Full article ">Figure 3
    <p>Analysis of inoculated Etrog citron plants by sPAGE and silver staining showing that they were infected with single viroids (1–5). CVd-II was not homogeneous but conformed by several bands (<bold>A</bold>,<bold>C</bold>) with slightly distinct electrophoretic mobilities (<bold>B</bold>) that were further characterized as specific variants of <italic>Hop stunt viroid</italic> (HSVd) showing faster mobility than CEVd (<bold>D</bold>).</p>
    Full article ">Figure 4
    <p>Characteristic symptoms as a result of viroid infection in plants grafted on trifoliate orange: Bark scaling induced by CEVd in trifoliate orange (<bold>A</bold>); bark cracking induced by <italic>Citrus bark cracking viroid</italic> (CBCVd) in trifoliate orange (<bold>B</bold>); wood pitting and gum deposits in the clementine scion caused by cachexia strains of HSVd (<bold>C</bold>); green streaks under the cracks induced by CBCVd in trifoliate orange (<bold>D</bold>). Lesions in the roots caused by CEVd in Citrange carrizo roots (<bold>E</bold>,<bold>F</bold>).</p>
    Full article ">Figure 5
    <p>The two scientists responsible for the early discovery/characterization of viroids as disease-causing agents and their conflictive publication.</p>
    Full article ">Figure 6
    <p>The dedication to cooperation issues in different parts of the world showed that in certain countries, as in Bhutan, the philosophy of life is to be taken into consideration.</p>
    Full article ">

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