Detection of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> in Patient and Mouse Blood and Ticks by a Duplex Real-Time PCR Assay

<div><p>Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, zoonotic infectious diseases caused by <i>Anaplasma phagocytophilum</i> and <i>Ehrlichia chaffeensis</i>, respectively. Early diagnosis is essential for rapid clinical treatment to avoid misdiagnosis and severe patient outcomes. Simple, sensitive and reliable diagnostic methods are urgently needed. In this study, we developed a duplex real-time PCR assay targeting the <i>A. phagocytophilum ankA</i> gene and the <i>E. chaffeensis</i> TRP120 gene, respectively. The lowest limit of detection of the duplex real-time PCR assay was 100 copies of the targeted <i>A. phagocytophilum ankA</i> gene and the <i>E. chaffeensis</i> TRP120 gene per reaction, and the specificity was 100%. Detection in blood DNA samples from the acute stage of illness for 22 HGA cases and 8 HME cases indicated that the duplex real-time PCR assay was more sensitive than the nested PCR assay. The infection of <i>Citellus</i><i>undulatus</i><i> Pallas</i> with <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> was first confirmed in Xinjiang Province and the positive rate was 3.1% for <i>A. phagocytophilum</i>, 6.3% for <i>E. chaffeensis</i> and 3.1% for co-infection with both pathogens. The rates of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> infection of <i>D</i><i>. silvarum</i> ticks collected from Shanxi Province were 8.2% and 14.8%, respectively, and the co-infection rate was 3.3%. The rates of <i>A. phagocytophilum</i> and <i>E. chaffeensis</i> infection in <i>H. longicornis</i> ticks collected from Shandong Province were 1.6% and 6.3%, respectively, and the co-infection rate was 1.6%.</p> </div

Molecular Characterization of <i>msp2/p44</i> of <i>Anaplasma phagocytophilum</i> Isolated from Infected Patients and <i>Haemaphysalis longicornis</i> in Laizhou Bay, Shandong Province, China

<div><p>Molecular characterization of the MSP2/P44 protein of <i>Anaplasma phagocytophilum</i> may determine not only if the bacterium is capable of invading hosts but also whether it generates antigenic variation for the purpose of escaping the host immune response, resulting in various pathologic injuries and serious clinical outcomes. Chinese anaplasmosis patients usually present with serious manifestations, and the fatality rate is as high as 26.5%. In this study, we amplified, cloned and sequenced the <i>msp2/p44</i> genes of three Chinese <i>A. phagocytophilum</i> isolates from Laizhou Bay, Shandong Province, where human granulocytic anaplasmosis (HGA) patients present severe clinical manifestations, and analyzed their genetic characterization and structural features. We also compared them with the HZ and Webster <i>A. phagocytophilum</i> strains. The sequences for both strains are available in GenBank. Analyses indicated that Chinese <i>A. phagocytophilum</i> isolates were significantly different from the HZ and Webster strains in terms of nucleotide sequences, amino acid sequences and protein secondary and tertiary structures. Moreover, the number of immunologic B-cell epitopes (19) of the MSP2 protein of the Chinese isolates was higher than that of the <i>A. phagocytophilum</i> strains HZ (16) and Webster (9). This genetic diversity of the MSP2/P44 protein of Chinese <i>A. phagocytophilum</i> isolates might be relevant and might have serious clinical outcomes. This observation could provide a clue to further understand the pathogenesis of Chinese <i>A. phagocytophilum</i>. </p> </div

Phylogenetic tree based on the <i>msp2/p44</i> nucleotide sequences (a) and MSP2/P44 amino acid sequences (b) generated using the neighbor-joining method.

<p><b>a</b>. Bootstrap values (>60%) are shown next to the nodes of the tree, and the scale bar indicates the number of nucleotide substitutions per site; <b>b</b>. The MSP2/P44 amino acid sequences were obtained by <i>msp2/p44</i> gene sequence translation, and the bootstrap values (>50%) are shown next to the nodes of the tree. The scale bar indicates the number of amino acid substitutions per site. The Chinese isolate LZ-HGA-Agent (red) and the international reference strains for APH-Webster and APH-HZ (blue) are highlighted. APH: <i>Anaplasma phagocytophilum</i>.</p

Copolymer-Functionalized Cellulose Nanocrystals as a pH- and NIR-Triggered Drug Carrier for Simultaneous Photothermal Therapy and Chemotherapy of Cancer Cells

As a class of biocompatible and biodegradable naturally derived nanomaterials, cellulose nanocrystals (CNCs) with diverse surface functionalization have aroused considerable attention for a range of biomedical applications in drug or gene delivery, as a fluorescent nanoprobe, in cancer targeting, and in photothermal cancer therapy, among others. Herein, we construct the copolymer-functionalized CNCs as a pH- and near-infrared (NIR)-triggered drug carrier for simultaneous photothermal therapy and chemotherapy of cancer cells. Poly(ε-caprolactone)-b-poly(2-(dimethylamino)ethyl methacrylate) (PCL-b-PDMAEMA) was conjugated onto the surface of CNCs through ring-opening polymerization, followed by activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP). The resultant CNC-based drug carrier can encapsulate doxorubicin (DOX) as a therapeutic agent and indocyanine green (ICG) as an NIR dye in the PCL core and the PDMAEMA shell, respectively, via hydrophobic and electrostatic interactions. In addition to the intrinsic pH response, the release profile of DOX can also be controlled by the duration of laser irradiation due to collapse of the crystal structure of the PCL domain with the increase of temperature induced by photothermal conversion. The drug carrier can exhibit enhanced cytotoxicity toward HepG2, human hepatocyte carcinoma, cells upon laser irradiation, which can be attributed to the synergistic effect arising from NIR-triggered burst release of DOX and photothermal heating. The rod-like morphology of the CNC-based drug carrier may help accelerate the endocytosis in cell membranes compared with its common spherical counterpart. Based on the abovementioned advantages, copolymer-functionalized CNCs can serve as a promising candidate for effective cancer treatment

Detection of Oxygen Vacancies in Oxides by Defect-Dependent Cataluminescence

Oxygen vacancies can control a number of distinct properties of oxides. However, rapid and simple detection of oxygen vacancies is a great challenge owing to their elusive species and highly diluted contents. In this work, we have discovered that cataluminescence (CTL) intensity in diethyl ether oxidation reaction on the surface of TiO₂ nanoparticles is proportional to the content of oxygen vacancies. The oxygen vacancy-dependent diethyl ether CTL is attributed to the fact that abundant chemisorbed O₂ in oxygen vacancies could facilitate its contact reaction with chemisorbed diethyl ether molecules, resulting in an obvious improvement of CTL intensity. Therefore, diethyl ether CTL can be employed as a simple probe for oxygen vacancies in TiO₂ nanoparticles. Its feasibility is validated by detecting the CTL intensity of diethyl ether on the surface of TiO₂ with variable oxygen vacancies by metal ion-doped TiO₂ nanoparticles (Cu, Fe, Co, and Cr) and hydrogen-treated TiO₂ nanoparticles at different temperatures. The content of oxygen vacancies by the present CTL probe is in good agreement with that obtained by conventional X-ray photoelectron spectroscopy (XPS) technique. The superior properties of the developed CTL probe over already-developed methods include fast response, easy operation, low cost, long-term stability, and simple configuration. We believe that the oxygen vacancy-sensitive CTL probe has a great potential in distinguishing oxygen vacancies in oxides

Predicted transmembrane domain for LZ-HGA-Agent-MSP2 (A), APH-HZ-MSP2 (B) and APH-Webster-MSP2 (C).

<p>The red legend (transmembrane —), blue legend (inside —) and pink legend (outside —) in panels A, B and C indicate the transmembrane domain, interior domain and exterior domain, respectively, for the MSP2 protein as predicted by the TMHMM program. The number on the horizontal abscissa in panels A, B and C indicates the amino acid (AA) residual site and size.</p

Aggregation-Induced Emission: A Simple Strategy to Improve Chemiluminescence Resonance Energy Transfer

The emergence of aggregation-induced emission (AIE) has opened up a new avenue for scientists. There is a great demand for the development of a new generation chemiluminescence resonance energy transfer (CRET) acceptors with AIE characteristics due to the aggregation-caused chemiluminescence (CL) quenching effect commonly observed in the conventional fluorophore CL acceptors at high concentrations. However, the systematical studies involving in AIE-amplified CL are still scarce. Herein, it is the first report that the gold nanocluster aggregates (a type of well-defined AIE molecules) are used to study their influence on the bis­(2,4,6-trichlorophenyl) oxalate (TCPO)–H₂O₂ CL reaction. Interestingly, the AIE molecules in the diluted solution are unable to boost the CL signal of the TCPO–H₂O₂ system, but their aggregates display a strongly enhanced CL emission compared to their counterparts of fluorophore molecules, thanks to the unique AIE effect of gold nanoclusters. In comparison to rhodamine B with the aid of an imidazole catalyst, the detection limit of the gold nanocluster aggregate-amplified CL probe for H₂O₂ (S/N = 3) is low in the absence of any catalyst. Finally, the other two typical AIE molecules, Au­(I)–thiolate complexes and 9,10-bis­[4-(3-sulfonatopropoxyl)-styryl]­anthracene (BSPSA), are investigated to verify the generality of the AIE molecule-amplified CL emissions. These results demonstrate effective access to highly fluorescent AIE molecules with practical applications in avoiding the aggregation-induced CL quenching at high concentrations, which can be expected to provide a novel and sensitive platform for the CL amplified detection

Classification results of normal elderly controls vs AD.

<p>Classification results of normal elderly controls vs AD.</p

Prediction and classification of Alzheimer disease based on quantification of MRI deformation - Fig 3

<p>(a) Classification sensitivity (green), specificity (blue), and accuracy (red) of normal elderly controls versus AD patients with different ROIs. The highest accuracy (96.5%) was achieved using the whole-brain gray matter as ROI with 93.85% sensitivity and 97.78% specificity. The algorithm obtained high sensitivity and specificity (>90%) with half of the ROIs. (b) The ROC curve of the prediction accuracy between normal controls versus AD. The AUCs were larger than 0.98 for the whole-brain gray matter and white matter (left), amygdala and hippocampus (middle), parietal and temporal lobes (right).</p

Improved Chemiluminescence in Fenton-Like Reaction via Dodecylbenzene-Sulfonate-Intercalated Layered Double Hydroxides

The interlamellar dodecylbenzene sulfonate (DBS) in layered double hydroxides (LDHs) can amplify an ultraweak chemiluminescence (CL) from Fenton-like reaction (Co­(II) + H₂O₂ + OH^–). The CL enhancement mechanism of the intercalated DBS on the H₂O₂–Co­(II) CL was investigated by CL spectrum, radical scavengers, powder X-ray diffraction measurements, scanning electron microscope images, transmission electron microscopy images, Fourier transform infrared spectroscopy, electronic spinning resonance measurements, and elemental analysis techniques. The hydrophobic microenvironment of the intercalated DBS in LDHs facilitated the formation of hydroxyl radical (•OH) by the reaction of Co­(II) with H₂O₂ in alkaline medium. The abundant •OH radical can easily react with the intercalated DBS in LDHs to form sulfite radical (•SO₃^–). The recombination of •SO₃^– radical could generate an intermediate as the excited sulfur dioxide molecules (SO₂*), which generated strong CL emission centered at ∼430 nm. Our experiments illustrated a powerful use of surfactant-intercalated LDHs in CL studies and could have potential applications in the quantitative determination of radical production