The complete chloroplast genome of the medicinally important plant <i>Plumbago zeylanica</i> L. (plumbaginaceae) and phylogenetic analysis

Plumbago zeylanica L. 1753 is a medicinally-important herb in family Plumbaginaceae. In this study, we assembled and reported the complete chloroplast genome of P. zeylanica. The plastome of P. zeylanica was 169,178 bp, including a large single-copy region of 92,135 bp, a small single-copy region (SSC) of 13,455 bp and a pair of inverted repeat regions (IRs) of 31,794 bp. It contained 124 genes, including 79 protein-coding genes, 37 tRNA genes and eight rRNA genes. Phylogenetic analysis showed that P. zeylanica formed a close relationship with P. auriculata in Plumbago. The first complete chloroplast genome report of P. zeylanica providing an opportunity to explore the genetic diversity, and would be also helpful in the species identification and conservation.</p

l‑Rhamnose Enhances the Immunogenicity of Melanoma-Associated Antigen A3 for Stimulating Antitumor Immune Responses

Vaccines based on melanoma-associated antigens (MAGEs) present a promising strategy for tumor immunotherapy, albeit with weak immunogenicity. In this study, the xenoantigen l-rhamnose (Rha) was chemically conjugated with truncated MAGE-A3 (tMAGE-A3) to generate Rha-tMAGE-A3. The product showed good antigenicity with anti-Rha antibodies purified from human serum. FITC-labeled Rha-tMAGE-A3 was detected in THP-1 human macrophage cells via the anti-Rha antibody-dependent antigen uptake process. Furthermore, peripheral blood mononuclear cells (PBMCs) stimulated with Rha-tMAGE-A3 in the presence of anti-Rha antibodies showed better cytotoxicity toward A375 human melanoma cells surfaced by MAGE-A3 antigen compared to PBMCs stimulated with tMAGE-A3. All data reveal that linking of Rha epitopes to MAGE enhances the immunogenicity of MAGE by harnessing the immune effector functions of human naturally existing anti-Rha antibodies. Rha epitopes could become immunogenicity enhancers of tumor associated antigens in the development of tumor immunotherapies

Dual-Emissive Phosphorescent Polymer Probe for Accurate Temperature Sensing in Living Cells and Zebrafish Using Ratiometric and Phosphorescence Lifetime Imaging Microscopy

Temperature plays an important part in many biochemical processes. Accurate diagnosis and proper treatment usually depend on precise measurement of temperature. In this work, a dual-emissive phosphorescent polymer temperature probe, composed of iridium­(III) complexes as temperature sensitive unit with phosphorescence lifetime of ∼500 ns and europium­(III) complexes as reference unit with lifetime of ∼400 μs, has been rationally designed and synthesized. Upon the increase of the temperature, the luminescence intensity from the iridium­(III) complexes is enhanced, while that from the europium­(III) complexes remains unchanged, which makes it possible for the ratiometric detection of temperature. Furthermore, the polymer also displays a significant change in emission lifetime accompanied by the temperature variation. By utilizing the laser scanning confocal microscope and time-resolved luminescence imaging systems, ratiometric and time-resolved luminescence imaging in Hela cells and zebrafish have been carried out. Notably, the intensity ratio and long-lifetime-based imaging can offer higher sensitivity, decrease the detection limit, and minimize the background interference from biosamples

Glycosylation of MBP with <i>E. coli</i> O157:H7 O-Ag and analyzed by (A), coomassie blue staining and western blot with (B), anti-6×-His antibody; (C), anti-MBP antibody; (D), O157:H7 antiserum.

<p>Glycosylation of MBP with <i>E. coli</i> O157:H7 O-Ag and analyzed by (A), coomassie blue staining and western blot with (B), anti-6×-His antibody; (C), anti-MBP antibody; (D), O157:H7 antiserum.</p

Antibody responses after third immunization.

<p>(<b>A</b>), <i>E. coli</i> O157:H7-specific IgG antibody titers of serum samples; (<b>B</b>), <i>E. coli</i> O157:H7-specific IgM antibody titers of serum samples; (<b>C</b>), <i>E. coli</i> O157:H7-specific IgA antibody titers of feces samples; (<b>D</b>), MBP-specific IgG antibody titers of serum samples. The cutoff value was OD_{negative control}×2.1. Results were expressed as the arithmetic mean ±SD indicated by error bars. Differences of two groups were indicated with symbols (*: P<0.05; **: P<0.01; ***: P<0.001).</p

Induction of Th1-biased responses by O-Ag-MBP.

<p>(<b>A</b>), Secretion of IFN-γ and IL-4 in serum samples of immunized mice. Differences of two groups are indicated with symbols (*: P<0.05; **: P<0.01). (<b>B</b>), <i>E. coli</i> O157:H7-specific IgG subtypes induced by O-Ag-MBP. BALB/c mice were immunized with O-Ag-MBP and serum samples were collected and <i>E. coli</i> O157:H7-specific IgG, IgG1, IgG2a and IgG2b titers were determined by ELISA. The cutoff value was OD_{negative control}×2.1. Results were expressed as the arithmetic mean ±SD indicated by error bars.</p

Spleen lymphocyte subsets assay.

<p>Spleen lymphocytes were isolated and stained for FITC-CD3, PE-CD4 or PE-CD8 antibodies and then analyzed by flow cytometry. The percentage of (<b>A</b>), CD3⁺ CD4⁺ T cells; (<b>B</b>), CD3⁺ CD8⁺ were expressed as the arithmetic mean ±SD indicated by error bars. Exprements were repeated three times. Differences of two groups are indicated with symbols (*: P<0.05; **: P<0.01).</p

Serum bactericidal activity against <i>E. coli</i> O157:H7.

<p>The % killing of <i>E. coli</i> O157:H7 are expressed as the arithmetic mean ±SD indicated by error bars.</p

Cytokines determined by ELISPOT.

<p>(<b>A</b>), IFN-γ; (<b>B</b>), IL-4. The spots for cytokine-producing lymphocytes after stimulation with MBP or O-Ag-MBP were counted and expressed based on 1×10⁵ cells. Results are expressed as the arithmetic mean ±SD indicated by error bars. Differences of three groups stimulated at the same dose of MBP are indicated with symbols (*: P<0.05; **: P<0.001).</p

MALDI-TOF MS analysis of (A) MBP and (B) O-Ag-MBP.

<p>(<b>A</b>), The peak with m/z 44017.39 corresponds to un-glycosylated MBP; (<b>B</b>), The peak with m/z 50530.62 and m/z 76617.71 correspond to glycosylated MBP (O-Ag-MBP).</p