Table_1_Differential Synergistic Interactions Among Four Different Wheat-Infecting Viruses.DOCX

Field-grown wheat (Triticum aestivum L.) plants can be co-infected by multiple viruses, including wheat streak mosaic virus (WSMV), Triticum mosaic virus (TriMV), brome mosaic virus (BMV), and barley stripe mosaic virus (BSMV). These viruses belong to four different genera in three different families and are, hence, genetically divergent. However, the impact of potential co-infections with two, three, or all four of them on the viruses themselves, as well as the wheat host, has yet to be examined. This study examined bi-, tri-, and quadripartite interactions among these viruses in wheat for disease development and accumulation of viral genomic RNAs, in comparison with single virus infections. Co-infection of wheat by BMV and BSMV resulted in BMV-like symptoms with a drastic reduction in BSMV genomic RNA copies and coat protein accumulation, suggesting an antagonism-like effect exerted by BMV toward BSMV. However, co-infection of either BMV or BSMV with WSMV or TriMV led to more severe disease than singly infected wheat, but with a decrease or no significant change in titers of interacting viruses in the presence of BMV or BSMV, respectively. These results were in stark contrast with exacerbated disease phenotype accompanied with enhanced virus titers caused by WSMV and TriMV co-infection. Co-infection of wheat by WSMV, TriMV, and BMV or BSMV resulted in enhanced synergistic disease accompanied by increased accumulation of TriMV and BMV but not WSMV or BSMV. Quadripartite interactions in co-infected wheat by all four viruses resulted in very severe disease synergism, leading to the death of the most infected plants, but paradoxically, a drastic reduction in BSMV titer. Our results indicate that interactions among different viruses infecting the same plant host are more complex than previously thought, do not always entail increases in virus titers, and likely involve multiple mechanisms. These findings lay the foundation for additional mechanistic dissections of synergistic interactions among unrelated plant viruses.</p

Investigating the Influences of Random-Region Length on Aptamer Selection Efficiency Based on Capillary Electrophoresis–SELEX and High-Throughput Sequencing

For aptamer selection, the random-region length of an ssDNA library was generally taken in a relatively arbitrary fashion, which may lead to failure for unsuitable target binding. Herein, we coupled high-efficiency capillary electrophoresis (CE)–SELEX and high-throughput sequencing (HTS) to investigate the influences of random-region length. First, one round of selection against programmed cell death-ligand 1 (PD-L1) was performed using ssDNA libraries with random-region lengths of 15, 30, 40, and 60 nt, respectively. A good correlation was observed between candidates’ random-region lengths and dissociation constant (Kd), in which the longer sequences presented higher affinity, and the picked Seq 60–1 after one round notably presented a similar affinity toward a reported aptamer through eight rounds. Molecular dynamics (MD) simulation suggested, for PD-L1, the long sequence could supply more noncovalent bonds including hydrogen bonds, electrostatic interactions, and hydrophobic interactions to form a stable protein/aptamer complex. Besides, four other proteins with selective binding performances validated the importance of random-region length. To further investigate how random-region length affects the selection efficiency, a mixed library with random-region lengths ranging from 10 to 50 nt was employed for six rounds of selection against Piezo2. Sequence variations were tracked by HTS, showing the preferential evolution and PCR uncertainty with even higher impact were the main causes. This study suggested random-region length plays a crucial factor, and a mixed library with different random-region sequences can be a worthy choice for increasing the speed of high-affinity aptamer selection. Moreover, the PCR process should be given particular attention in aptamer selection

Crystallization in Ionic Liquids: Synthesis, Properties, and Polymorphs of Uranyl Salts

Crystallizations in uranyl-containing ionic liquids yielding crystals {X}_a{[UO₂]_b[Y]_c} with selected anions and cations (X = [Bmim]⁺ for Y = Cl^–, NO₃^–, and SCN^–; Y = Cl^– for X = [Emim]⁺, [Emmim]⁺, [Bmim]⁺, and [Bmmim]⁺; Emim = l-ethyl-3-methyl-imidazolium, Emmim = l-ethyl-2,3-dimethylimidazolium, Bmim = l-butyl-3-methylimidazolium, and Bmmim = l-butyl-2,3-dimethylimidazolium) were performed herein. Through standard crystallographic analyses, Hirshfeld surface analyses, and multiple characterization techniques, compounds with common cations/anions were investigated. For compounds [Bmim]₂[(UO₂)₂­(μ-OH)₂(NO₃)₄] (<b>1</b>), [Bmim]₃­[UO₂(NCS)₅] (<b>2</b>), and [Bmim]₂­[UO₂Cl₄] (<b>3TT</b> and <b>3TG</b>), common [Bmim]⁺ cations with different conformations were studied with respect to packing and interactions (for <b>1</b>). The coordinated [(UO₂)₂(μ-OH)₂­(NO₃)₄]^2–, [UO₂(NCS)₅]^3–, and [UO₂Cl₄]^2– anions that have historically been related to nuclear fuel cycles were demonstrated with respect to geometry and distortion. For compounds with common [UO₂Cl₄]^2– anions [Emim]₂­[UO₂Cl₄] (<b>4</b>), [Emmim]₂­[UO₂Cl₄] (<b>5</b>), [Bmmim]₂­[UO₂Cl₄] (<b>6</b>), <b>3TT</b>, and <b>3TG</b>, observed interionic interactions that have been previously impeded by limited structural information were discussed fully in relation to different cations and temperatures. Moreover, multistep phase transformations of <b>2</b>, which have been undefined in solution studies, have been identified through differential scanning calorimetry analyses and polarizing optical microscopy. The polymorph transformations between <b>3TT</b> and <b>3TG </b> in solution, as controlled by uranyl concentration, were studied using optical microscopy and powder X-ray diffraction. The thermal stability, IR/Raman, and UV–vis/luminescence spectra of these compounds were also investigated

Straightforward and Ultrastable Surface Modification of Microfluidic Chips with Norepinephrine Bitartrate Improves Performance in Immunoassays

Polymers are commonly used materials for microfluidic chip fabrication, because they are standardized in fabrication and low in cost. However, most polymeric materials that are readily fabricated on the industrial scale are hydrophobic, which is inconvenient for the injection and flow of the aqueous solution, resulting in poor analytical performance for biochemical assays. In this work, we present a straightforward and ultrastable surface modification process for polymeric chips. A one-step modification by using norepinephrine bitartrate monohydrate as a modification reagent is completed at room temperature. The hydrophilicity of the polymeric surfaces increases dramatically. Surface modification is stable for at least 2.5 years, allowing for autoinjection of aqueous solution into the channels. The chips are applied in the immunoassay of alpha-fetoprotein (AFP). The low nonspecific adsorption after modification results in significantly decreased background noise, optimized signal-to-noise ratios (SNR), and dramatically enhanced reproducibility of the immunoassay. Thirty clinical human serum samples are analyzed; these results strongly correlated with the values obtained using commercial test kits. We anticipate that this surface modification method can be used for immunoassay devices in analytical and biosensing technology

Investigating the Influences of Random-Region Length on Aptamer Selection Efficiency Based on Capillary Electrophoresis–SELEX and High-Throughput Sequencing

For aptamer selection, the random-region length of an ssDNA library was generally taken in a relatively arbitrary fashion, which may lead to failure for unsuitable target binding. Herein, we coupled high-efficiency capillary electrophoresis (CE)–SELEX and high-throughput sequencing (HTS) to investigate the influences of random-region length. First, one round of selection against programmed cell death-ligand 1 (PD-L1) was performed using ssDNA libraries with random-region lengths of 15, 30, 40, and 60 nt, respectively. A good correlation was observed between candidates’ random-region lengths and dissociation constant (Kd), in which the longer sequences presented higher affinity, and the picked Seq 60–1 after one round notably presented a similar affinity toward a reported aptamer through eight rounds. Molecular dynamics (MD) simulation suggested, for PD-L1, the long sequence could supply more noncovalent bonds including hydrogen bonds, electrostatic interactions, and hydrophobic interactions to form a stable protein/aptamer complex. Besides, four other proteins with selective binding performances validated the importance of random-region length. To further investigate how random-region length affects the selection efficiency, a mixed library with random-region lengths ranging from 10 to 50 nt was employed for six rounds of selection against Piezo2. Sequence variations were tracked by HTS, showing the preferential evolution and PCR uncertainty with even higher impact were the main causes. This study suggested random-region length plays a crucial factor, and a mixed library with different random-region sequences can be a worthy choice for increasing the speed of high-affinity aptamer selection. Moreover, the PCR process should be given particular attention in aptamer selection

Straightforward and Ultrastable Surface Modification of Microfluidic Chips with Norepinephrine Bitartrate Improves Performance in Immunoassays

Polymers are commonly used materials for microfluidic chip fabrication, because they are standardized in fabrication and low in cost. However, most polymeric materials that are readily fabricated on the industrial scale are hydrophobic, which is inconvenient for the injection and flow of the aqueous solution, resulting in poor analytical performance for biochemical assays. In this work, we present a straightforward and ultrastable surface modification process for polymeric chips. A one-step modification by using norepinephrine bitartrate monohydrate as a modification reagent is completed at room temperature. The hydrophilicity of the polymeric surfaces increases dramatically. Surface modification is stable for at least 2.5 years, allowing for autoinjection of aqueous solution into the channels. The chips are applied in the immunoassay of alpha-fetoprotein (AFP). The low nonspecific adsorption after modification results in significantly decreased background noise, optimized signal-to-noise ratios (SNR), and dramatically enhanced reproducibility of the immunoassay. Thirty clinical human serum samples are analyzed; these results strongly correlated with the values obtained using commercial test kits. We anticipate that this surface modification method can be used for immunoassay devices in analytical and biosensing technology

Correlation of sphingolipid biomarkers with ear swelling and splenic index.

<p>Correlation of sphingolipid biomarkers with ear swelling and splenic index.</p

Visualization of sphingolipid metabolites in the plasma, kidney, spleen and liver.

<p>The figure depicts sphingolipid metabolites that participate in the early steps of sphingolipid biosynthesis. The pathway maps are overlaid by the fold changes in subclasses of sphingolipid metabolites between the control group and other groups by asterisks indicating the statistical significance on the right side (* = P<0.05, ** = P<0.01; n = 8). The content of each sphingolipid metabolite was determined as the mean of eight independent parallel samples.</p

Sphingolipids as New Biomarkers for Assessment of Delayed-Type Hypersensitivity and Response to Triptolide

<div><h3>Background</h3><p>Hypersensitivity diseases are associated with many severe human illnesses, including leprosy and tuberculosis. Emerging evidence suggests that the pathogenesis and pathological mechanisms of treating these diseases may be attributable to sphingolipid metabolism.</p> <h3>Methods</h3><p>High performance liquid chromatography-tandem mass spectrometry was employed to target and measure 43 core sphingolipids in the plasma, kidneys, livers and spleens of BALB/c mice from four experimental groups: control, delayed-type hypersensitivity (DTH) model, DTH+triptolide, and control+triptolide. Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify potential biomarkers associated with variance between groups. Relationships between the identified biomarkers and disease markers were evaluated by Spearman correlation.</p> <h3>Results</h3><p>As a treatment to hypersensitivity disease, triptolide significantly inhibit the ear swelling and recover the reduction of splenic index caused by DTH. The sphingolipidomic result revealed marked alterations in sphingolipid levels between groups that were associated with the effects of the disease and triptolide treatment. Based on this data, 23 potential biomarkers were identified by OPLS-DA, and seven of these biomarkers correlated markedly with the disease markers (p<0.05) by Spearman correlation.</p> <h3>Conclusions</h3><p>These data indicate that differences in sphingolipid levels in plasma and tissues are related to DTH and treatment with triptolide. Restoration of proper sphingolipid levels may attribute to the therapeutic effect of triptolide treatment. Furthermore, these findings demonstrate that targeted sphingolipidomic analysis followed by multivariate analysis presents a novel strategy for the identification of biomarkers in biological samples.</p> </div

Sphingolipid composition of mice kidney or plasma measured by triple quadruples MS/MS.

<p>Four groups including: control, model, model+triptolide and control+triptolide, each of which contains 8 samples. Sphingolipids were isolated from kidney homogenate corresponding to 1 mg protein or from 0.1 mL plasma. Bars are expressed as means ± SD, values for each sample are the average of 8 samples separately (pmol/mg protein, 0.1 mL plasma). Statistical difference from control or model group is indicated with an asterisk or a “&”, respectively. * or &: p<0.05 and ** or &&: p<0.01.</p