Increased spleen weight and T cell activation in <i>Tlr9</i>^<i>-/-</i> MRL/+ mice.

<p><b>(A)</b> Spleens were weighed. <b>(B)</b> Ly6G⁺CD11b⁺ neutrophils and <b>(C)</b> CD19^-CD11c⁺I-A/I-E⁺ dendritic cells were evaluated by FACS. <b>(D-E)</b> Naive (CD44^-CD62L⁺) cells were evaluated by FACS among TCRß⁺CD4⁺ (B) and TCRß⁺CD8⁺ (C) populations. * p<0.05; ** p<0.01; **** p<0.0001 by two-tailed Mann-Whitney U-test. Data are pooled from five (A) or four (B-E) experimental cohorts.</p

Autoantibody production in <i>Tlr9</i>^<i>-/-</i> MRL/+ mice.

<p><b>(A)</b> Serum anti-nucleosome IgG autoantibodies were measured by ELISA and are expressed relative to a PL2-3 standard. <b>(B)</b> Serum anti-Sm IgG autoantibodies were measured by ELISA and are expressed relative to a Y2 standard. <b>(C)</b> Serum anti-RNA IgG autoantibodies were measured by ELISA and are expressed relative to a BWR4 standard. <b>(D)</b> Serum kappa anti-IgG2a rheumatoid factor autoantibodies were measured by ELISA and are expressed relative to a 400tμ23 standard. * p<0.05; *** p<0.001; **** p<0.0001 by two-tailed Mann-Whitney U-test.</p

Toll-like receptor 9 suppresses lupus disease in <i>Fas</i>-sufficient MRL Mice

<div><p>Genetic deficiency in TLR9 accelerates pathogenesis in the spontaneous polygenic MRL.<i>Fas</i>^<i>lpr</i> murine model of systemic lupus erythematosus, despite the absence of anti-nucleosome autoantibodies. However, it could be argued that this result was dependent on <i>Fas</i>-deficiency rather than lupus-promoting genes in the MRL genetic background. Here we report the effects of TLR9 deficiency on autoimmune disease independent of the <i>lpr</i> mutation in <i>Fas</i> by characterizing <i>Tlr9</i>^<i>-/-</i> and <i>Tlr9</i>^<i>+/+</i> mice on the <i>Fas</i>-intact MRL/+ genetic background. By 30 weeks of age, <i>Tlr9-</i>deficient MRL/+ had more severe renal disease, increased T cell activation, and higher titers of anti-Sm and anti-RNA autoantibodies than <i>Tlr9</i>-intact animals, as had been the case in the MRL.<i>Fas</i>^<i>lpr</i> model. In addition, <i>Tlr9</i>-deficient MRL/+ mice had increased numbers of germinal center phenotype B cells and an increase in splenic neutrophils and conventional dendritic cell populations. Thus, the disease accelerating effects of <i>Tlr9</i> deficiency are separable from those mediated by the <i>Fas</i> mutation in the lupus-prone MRL genetic background. Nonetheless, disease acceleration in <i>Tlr9</i>-deficient MRL/+ mice was phenotypically distinct from that in <i>Fas</i>-deficient counterparts, which has important implications.</p></div

Renal disease is suppressed by <i>Tlr9</i> in MRL/+ mice.

<p><b>(A)</b> Proteinuria was evaluated in 30 week old mice of the indicated genotypes by dipstick assay. <b>(B)</b> Severity of glomerular disease on H&E stained sections was evaluated by a pathologist on a 0–6 scale. <b>(C)</b> Perivascular and interstitial renal infiltrates were evaluated by a pathologist on a 0–4 scale. <b>(D)</b> Skin lesions were scored based on area with up to an additional 0.5 points for facial rash / loss of whiskers and 0.25 points for dermatitis of each ear. In all graphs, horizontal lines represent medians and each point represents an individual animal. ** p<0.01; **** p<0.0001 by two-tailed Mann-Whitney U-test. Data are pooled from five experimental cohorts.</p

HEp-2 antinuclear antibody staining patterns are changed in <i>Tlr9</i>^<i>-/-</i> MRL/+ mice.

<p><b>(A)</b> Representative HEp-2 staining patterns from <i>Tlr9</i>^<i>+/+</i> MRL/+ (<i>top</i>) and <i>Tlr9</i>^<i>-/-</i> MRL/+ (<i>bottom</i>) serum. <b>(B)</b> Proportion of sera with the indicated dominant staining pattern from <i>Tlr9</i>^<i>+/+</i> MRL/+ (<i>left</i>) or <i>Tlr9</i>^<i>-/-</i> MRL/+ (<i>right</i>). <b>(C)</b> Percentage of sera of indicated genotypes which stained positive for mitotic bodies in the HEp-2 ANA assay. **** p<0.0001 by two-tailed Fisher's exact test. <b>(D)</b> Relative intensity of cytoplasmic staining in the HEp-2 ANA assay. * p<0.05 by two-tailed Mann-Whitney U-test.</p

Efficient Sorption and Removal of Perfluoroalkyl Acids (PFAAs) from Aqueous Solution by Metal Hydroxides Generated in Situ by Electrocoagulation

Removal of environmentally persistent perfluoroalkyl acids (PFAAs), that is, perfluorooctanesulfonate (PFOS) and perfluorocarboxylic acids (PFCAs, C₄ ∼ C₁₀) were investigated through sorption on four metal hydroxide flocs generated in situ by electrocoagulation in deionized water with 10 mM NaCl as supporting electrolyte. The results indicated that the zinc hydroxide flocs yielded the highest removal efficiency with a wide range concentration of PFOA/PFOS (1.5 μM ∼ 0.5 mM) at the zinc dosage <150 mg L^–1 with the energy consumption <0.18 Wh L^–1. The sorption kinetics indicated that the zinc hydroxide flocs had an equilibrium adsorbed amount (<i>q</i>_e) up to 5.74/7.69 mmol g^–1 (Zn) for PFOA/PFOS at the initial concentration of 0.5 mM with an initial sorption rate (<i>v</i>₀) of 1.01 × 10³/1.81 × 10³ mmol g^–1 h^–1. The sorption of PFOA/PFOS reached equilibrium within <10 min. The sorption mechanisms of PFAAs on the zinc hydroxide flocs were proposed based on the investigation of various driving forces. The results indicated that the hydrophobic interaction was primarily responsible for the PFAAs sorption. The electrocoagulation process with zinc anode may have a great potential for removing PFAAs from industrial wastewater as well as contaminated environmental waterbody

Influence of the Net Charge on the Reactivity of a Manganese(IV) Species: Leading to the Correlation of Its Physicochemical Properties with Reactivity

Clarifying how versatile physicochemical parameters of an active metal intermediate affect its reactivity would help to understand its roles in chemical and enzymatic oxidations. The influence of the net charge on electron transfer and hydrogen abstraction reactions of a manganese­(IV) species having hydroxide ligand has been investigated here. It was found that increasing one unit of the positive net charge from 2+ to 3+ would accelerate its electron-transfer rate by 10–20 fold in oxygenation of tris­(4-methoxyphenyl)­phosphine. In contrast, the hydrogen abstraction rate is insensitive to its net charge change, and the insensitivity has been attributed to the compensation effect between the redox potential and p<i>K</i>_a, which determine the hydrogen abstraction capability of a metal ion. Similar net-charge-promoted electron transfer but not hydrogen abstraction has also been observed in intramolecular electron transfer and hydrogen abstraction reactions when using thioxanthene as substrate. Together with the previous understanding of the reactivity of the identical manganese­(IV) species having Mn^IV–OH or Mn^IV=O functional groups, the relationships of the oxidative reactivity of an active metal intermediate with its physicochemical parameters such as the net charge, the redox potential and the metal–oxygen bond order (M–O versus MO) have been discussed with this manganese­(IV) model

Up-regulated expression of the ER chaperone GRP78 at the mRNA and protein levels in the different cataract lens groups RNA and protein were extracted from human lens specimens.

<p>Western blotting was performed to detect the protein expression of GRP78 (Fig 1A and 1B), and β-actin was used as the internal control; real-time PCR was performed to detect the gene expression level of GRP78 in each group (Fig 1C), and 18S rRNA was used as the internal control gene (mean ± SD, <i>n</i> = 3). *<i>P</i> < 0.05; **<i>P</i> < 0.001.</p

Up-regulated p-IRE1α protein expression level and spliced XBP1 gene expression level in the different cataract lens groups RNA and protein were extracted from human lens specimens.

<p>Western blotting was performed to detect the protein expression of p-IRE1α (Fig 2A, Fig 2B), and β-actin was used as the internal control; real-time PCR was performed to detect the relative gene expression levels of spliced XBP1 in each group (Fig 2C), and 18S rRNA was used as the internal control gene (mean ± SD, <i>n</i> = 3). *<i>P</i> < 0.05; **<i>P</i> < 0.001.</p

Up-regulated cleaved ATF6 protein and gene expression levels in the different cataract lens groups.

<p>RNA and protein were extracted from human lens specimens. Western blotting was performed to detect the protein expression of cleaved ATF6 (Fig 4A and 4B), and β-actin was used as the internal control; real-time PCR was performed to detect the relative gene expression level of ATF6 in each group (Fig 4C), and 18S rRNA was used as the internal control gene (mean ± SD, <i>n</i> = 3). *<i>P</i> < 0.05; **<i>P</i> < 0.001.</p