Chemical Degradation of Drinking Water Disinfection Byproducts by Millimeter-Sized Particles of Iron−Silicon and Magnesium−Aluminum Alloys

Chemical Degradation of Drinking Water Disinfection Byproducts by Millimeter-Sized Particles of Iron−Silicon and Magnesium−Aluminum Alloy

Histologic analysis of cellular infiltrations in the liver of TAM^−/− mice.

<p>(A) H&E staining of paraffin-embedded liver sections of TAM^−/− mice at the indicated ages. M1, M6 and M12 represent the liver sections of 1, 6 and 12 month-old mice respectively. Note a progression of cellular infiltrations in the liver as TAM^−/− mice aged. Arrows indicate focal infiltrates. (B) Characterization of infiltrated lymphocytes. Immunofluorescence was performed using FITC-conjugated antibodies against lymphocyte markers. Note that the infiltrated lymphocytes predominantly consist of CD4⁺ and CD8⁺ T cells. (C) Quantitatively analysis of lymphocytes. The cells were labeled with FITC-conjugated antibodies against CD4, CD8 and B220, and subsequently subject to flow cytometry. (D) Identification of macrophages. Immunohistochemistry was used for the identification of total macrophages (F4/80⁺), circulating macrophages (ED1⁺) and resident macrophages (ED2⁺). Note that the macrophages consist of resident and infiltrated cells in the liver of TAM^−/− mice, whereas only resident macrophages were observed in WT controls. Frozen liver sections of 10-month-old mice were used for the immune staining. Images are representatives of at least 5 mice. Scale bar  = 20 µm.</p

Expression of TAM RTKs, Gas6 and Protein S.

<p>(A) Western blot analyses of the liver lysates for the examination of TAM RTKs, Gas6 and Protein S. (B) Expression of TAM RTKs, Gas6 and Protein S in isolated liver cells: parenchymal cells (PCs), Kupffer cells(KCs) and sinusoidal endothelial cells (SECs). The primary cells were subjected to Western blotting. (C) Immunohistochemistry for the detection of TAM RTKs, Gas6 and Protein S. Arrowheads indicate PCs, and arrows indicate spindle-shaped sinusoidal cells corresponding to KCs and SECs. In negative controls (islets), sections were incubated with primary antibodies pre-incubated with an excess of blocking peptide. The livers of 15-week-old WT mice were used for the protein analyses. The images are representatives of at least three experiments. Scale bar  = 20 µm.</p

Autoantibodies diagnostic of AIH in sera from TAM^−/−<b>mice.</b>

<p>(A) Immunofluorescence on Hepa1-6 cells for detection of ANAs. The images are representative staining patterns using ×200 diluted sera of 12-month-old WT and TAM^−/− mice (n = 5 mice each genotype). Scale bar  = 40 µm. (B) The sera were diluted in series. Each circle indicates the maximum dilution of sera from individual mice, in which ANAs can be detectable. (C) Measurement of anti-actin autoantibody component of SMA using ELISA. Each circle indicates unit of anti-actin autoantibodies in sera of individual mice. (D) Titer of SMA. The microscope slides (Rat stomach) were stained using sera at the indicated dilutions. (E) Plasma IgG. The plasma were collected from 12-month-old mice. IgG levels were measured using ELISA.</p

Bone marrow transplantation.

<p>Lethally irradiated 6-month-old mice were injected with bone marrow from 2-month-old donor mice through a tail vein. The engrafted mice were analyzed at 6 months after bone marrow transplantation. (A) Expression of inflammatory cytokines in the liver of WT→TAM^−/− mice. (B) Activities of serum ALT and AST in WT→TAM^−/− mice. (C) Cytokine expression in the liver of TAM^−/−→WT mice. (D) Serum AST and ALT activities in the TAM^−/−→WT mice. Total RNA was extracted from the liver, and the mRNAs of cytokines were determined by real-time RT-PCR. Serum AST and ALT activities were measured by enzymatic assay kits. (E) The expression of Axl in the circulating leukocytes in engrafted mice. Data are mean ± SEM (n = 10). **<i>P</i><0.01.</p

Applications of Multiple Nuclear Genes to the Molecular Phylogeny, Population Genetics and Hybrid Identification in the Mangrove Genus <i>Rhizophora</i>

<div><p>The genus <i>Rhizophora</i> is one of the most important components of mangrove forests. It is an ideal system for studying biogeography, molecular evolution, population genetics, hybridization and conservation genetics of mangroves. However, there are no sufficient molecular markers to address these topics. Here, we developed 77 pairs of nuclear gene primers, which showed successful PCR amplifications across all five <i>Rhizophora</i> species and sequencing in <i>R</i>. <i>apiculata</i>. Here, we present three tentative applications using a subset of the developed nuclear genes to (I) reconstruct the phylogeny, (II) examine the genetic structure and (III) identify natural hybridization in <i>Rhizophora</i>. Phylogenetic analyses support the hypothesis that <i>Rhizophora</i> had disappeared in the Atlantic-East Pacific (AEP) region and was re-colonized from the IWP region approximately 12.7 Mya. Population genetics analyses in four natural populations of <i>R</i>. <i>apiculata</i> in Hainan, China, revealed extremely low genetic diversity, strong population differentiation and extensive admixture, suggesting that the Pleistocene glaciations, particularly the last glacial maximum, greatly influenced the population dynamics of <i>R</i>. <i>apiculata</i> in Hainan. We also verified the hybrid status of a morphologically intermediate individual between <i>R</i>. <i>apiculata</i> and <i>R</i>. <i>stylosa</i> in Hainan. Based on the sequences of five nuclear genes and one chloroplast intergenic spacer, this individual is likely to be an F1 hybrid, with <i>R</i>. <i>stylosa</i> as its maternal parent. The nuclear gene markers developed in this study should be of great value for characterizing the hybridization and introgression patterns in other cases of this genus and testing the role of natural selection using population genomics approaches.</p></div

Liver damage in TAM^−/− mice.

<p>(A) Representative images of H&E stained paraffin-embedded liver sections of 12-month-old WT and TAM^−/− female mice. Scale bar  = 10 µm. (B) Serum ALT and AST levels in 12-month-old mice. Each circle represents unit of ALT or AST in sera of an individual mouse. (C) Serum ALT and AST levels in mice at the indicated ages. Note that the livers were progressively damaged as TAM^−/− mice aged. (D) Serum ALT and AST levels in 12-month-old mice mutant singly, doubly, and triply for TAM RTKs. T, A, and M represent Tyro3, Axl and Mer, respectively. Data are mean ± SEM, n≥10. *<i>P</i><0.05, **<i>P</i><0.01.</p

Primers used for real-time RT-PCRs.

<p>Primers used for real-time RT-PCRs.</p

Activation of TLR signaling in the liver of TAM^−/− mice.

<p>(A) Expression of inflammatory cytokines. The mRNAs were analyzed by real-time RT-PCR (left panel) and the protein levels were determined by ELISA assay (right panel). Total RNAs were extracted from the liver and analyzed for the cytokine expression using RT-PCR, and cytokine levels in liver lysates were measured using ELISA. Data are mean ± SEM of three independent experiments. **<i>P</i><0.01. (B) Analysis on the phosphorylation of NF-κB and IRF3. The liver lysates were used for immunoblots probed for phosphor-P65 (p-P65) and phosphor-IRF3 (p-IRF3).</p